U.S. patent number 11,266,685 [Application Number 15/317,316] was granted by the patent office on 2022-03-08 for silicone-based biophotonic compositions and uses thereof.
This patent grant is currently assigned to KLOX TECHNOLOGIES INC.. The grantee listed for this patent is KLOX Technologies Inc.. Invention is credited to Abdellatif Chenite, Eric DesRosiers, Emmanuelle Devemy, Joanna Jaworska, Nikolaos Loupis, Remigio Piergallini.
United States Patent |
11,266,685 |
Piergallini , et
al. |
March 8, 2022 |
Silicone-based biophotonic compositions and uses thereof
Abstract
The present disclosure provides silicone-based biophotonic
compositions and methods useful in phototherapy. In particular, the
silicone-based biophotonic compositions of the present disclosure
include a silicone phase and a surfactant phase, wherein the
surfactant phase comprises at least one chromophore solubilized in
a surfactant. The silicone-based biophotonic compositions and the
methods of the present disclosure are useful for promoting wound
healing and scarring, as well as various other skin disorders.
Inventors: |
Piergallini; Remigio
(Grottammare, IT), Loupis; Nikolaos (Athens,
GR), Jaworska; Joanna (Montreal, CA),
Devemy; Emmanuelle (Montreal, CA), DesRosiers;
Eric (Outremont, CA), Chenite; Abdellatif
(Kirkland, CA) |
Applicant: |
Name |
City |
State |
Country |
Type |
KLOX Technologies Inc. |
Laval |
N/A |
CA |
|
|
Assignee: |
KLOX TECHNOLOGIES INC. (Laval,
CA)
|
Family
ID: |
1000006159053 |
Appl.
No.: |
15/317,316 |
Filed: |
June 9, 2015 |
PCT
Filed: |
June 09, 2015 |
PCT No.: |
PCT/IB2015/001761 |
371(c)(1),(2),(4) Date: |
December 08, 2016 |
PCT
Pub. No.: |
WO2015/189712 |
PCT
Pub. Date: |
December 17, 2015 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20170209484 A1 |
Jul 27, 2017 |
|
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
62009870 |
Jun 9, 2014 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
41/00 (20130101); A61K 8/416 (20130101); A61K
47/10 (20130101); A61Q 19/00 (20130101); A61K
9/0014 (20130101); A61K 8/891 (20130101); A61K
8/731 (20130101); A61L 26/0095 (20130101); A61Q
19/08 (20130101); A61N 5/062 (20130101); A61K
8/895 (20130101); A61K 31/352 (20130101); A61K
8/498 (20130101); A61K 47/20 (20130101); A61L
26/0061 (20130101); A61K 8/90 (20130101); A61K
41/008 (20130101); A61K 31/80 (20130101); A61K
8/86 (20130101); A61N 5/0616 (20130101); A61K
8/463 (20130101); A61L 26/0095 (20130101); C08L
83/04 (20130101); A61L 26/0095 (20130101); C08L
71/02 (20130101); A61N 2005/0662 (20130101); A61K
2800/81 (20130101); A61K 9/1075 (20130101); A61K
9/7015 (20130101); A61N 2005/0651 (20130101); A61K
2800/434 (20130101); A61K 2800/52 (20130101); A61K
2800/596 (20130101); A61K 47/34 (20130101); A61K
2800/48 (20130101) |
Current International
Class: |
A61K
31/80 (20060101); A61K 47/20 (20060101); A61Q
19/00 (20060101); A61K 8/49 (20060101); A61L
26/00 (20060101); A61Q 19/08 (20060101); A61K
8/895 (20060101); A61K 8/90 (20060101); A61K
8/73 (20060101); A61K 31/352 (20060101); A61N
5/06 (20060101); A61K 47/10 (20170101); A61K
41/00 (20200101); A61K 8/86 (20060101); A61K
8/891 (20060101); A61K 9/00 (20060101); A61K
8/46 (20060101); A61K 8/41 (20060101); A61K
47/34 (20170101); A61K 9/70 (20060101); A61K
9/107 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
|
|
|
|
|
2408323 |
|
Nov 2001 |
|
CA |
|
2902363 |
|
Sep 2014 |
|
CA |
|
2916337 |
|
Jan 2015 |
|
CA |
|
102300587 |
|
Dec 2011 |
|
CN |
|
102480969 |
|
May 2012 |
|
CN |
|
2499921 |
|
Sep 2013 |
|
GB |
|
H09505066 |
|
May 1997 |
|
JP |
|
H11506769 |
|
Jun 1999 |
|
JP |
|
2005-034353 |
|
Feb 2005 |
|
JP |
|
2005527493 |
|
Sep 2005 |
|
JP |
|
9513792 |
|
May 1995 |
|
WO |
|
9639116 |
|
Dec 1996 |
|
WO |
|
01/58452 |
|
Aug 2001 |
|
WO |
|
2003061696 |
|
Jul 2003 |
|
WO |
|
2010051636 |
|
May 2010 |
|
WO |
|
2011006100 |
|
Jan 2011 |
|
WO |
|
2011108520 |
|
Sep 2011 |
|
WO |
|
2001085212 |
|
Nov 2011 |
|
WO |
|
2012119131 |
|
Sep 2012 |
|
WO |
|
2013155620 |
|
Oct 2013 |
|
WO |
|
2014040176 |
|
Mar 2014 |
|
WO |
|
2014040177 |
|
Mar 2014 |
|
WO |
|
2014138930 |
|
Sep 2014 |
|
WO |
|
2015000058 |
|
Jan 2015 |
|
WO |
|
Other References
Brick et al., High tear strength silicone elastomers with low
hardness and high elongation. International SAMPE Technical
Conference, pp. 1-10. (Year: 2012). cited by examiner .
SYLGARD.RTM., "184 Silicone Elastomer Product Information Sheet",
[Online], Apr. 2, 2014, Retrieved from the Internet URL:
http://www.dowcomine.com/DataFiles/090276fe80190b08.pdf. cited by
applicant .
Curtis et al., "Medical applications of silicones", Biomaterials
Sciences, An Introduction to Materials in Medicine. cited by
applicant .
Database GNPD, [Online] MINTEL, Nov. 30, 2010, "Photo dynamic
therapy SPF 30", accession No. 1442681, XP002775115. cited by
applicant .
Supplementary European Search Report of European Patent Application
No. 15806945.0; dated Oct. 31, 2017; Munich; Olausson Boulois, J.
cited by applicant .
International Search Report of PCT/IB2015/001761, dated Jan. 28,
2016, Wesley Sharman. cited by applicant .
Office Action issued from the Japanese Patent Office dated Dec. 3,
2019. cited by applicant .
Office Action issued from the European Patent Office dated Dec. 10,
2019. cited by applicant .
English abstract provided for JP 2005-034353. cited by applicant
.
Avouac et al.. Inhibition of activator protein 1 signaling
abrogates transforming growth factor b-mediated activation of
fibroblasts and prevents experimental fibrosis, Arthritis
Rheumatism, 2012, vol. 64: 1642-1652. cited by applicant .
Beyer et al., Tyrosine kinase signaling in fibrotic disorders,
Translation of basic research to human disease, Biochem Biophys
Acta, 2013, vol. 1832: 897-904. cited by applicant .
Chen et al.. Focus on collagen: In vitro systems to study
fibrogenesis and antifibrosis--state of the art, Fibrogenesis
Tissue Repair, 2009, vol. 2: 7. cited by applicant .
Cutroneo, TGF-beta-induced fibrosis and SMAD signaling, oligo
decoys as natural therapeutics for inhibition of tissue and
scarring, Wound Rep Regen 2007, vol. 15, S54-60. cited by applicant
.
Gauglitz et al.. Hypertrophic scarring and keloids: pathomechanisms
and current and emerging treatment strategies, Mol Med, 2011, vol.
17:113-125. cited by applicant .
Momtazi et al., A nude mouse model of hypertophic scar shows
morphologic and histologic characteristics of human hypertrophic
scar, Wound Rep Reg, 2013, vol. 21: 77-87. cited by applicant .
Trojanowska, Role of PDGF in fibrotic diseases and systemic
sclerosis, Rheumatology, 2008, vol. 47, v2-v4. cited by
applicant.
|
Primary Examiner: Vanhorn; Abigail
Attorney, Agent or Firm: BCF LLP
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a national stage application under 35 U.S.C.
.sctn. 371 of International Application No. PCT/IB2015/001761,
filed Jun. 9, 2015, which claims the benefit of and priority to
U.S. Provisional Patent Application No. 62/009,870 filed Jun. 9,
2014, which is incorporated by reference herein in its entirety.
International Application PCT/IB2015/001761 was published under PCT
Article 21(2) in English.
Claims
The invention claimed is:
1. A silicone-based biophotonic membrane comprising a silicone
phase and a thermogellable surfactant phase, wherein the
thermogellable surfactant phase comprises at least one chromophore
solubilized in a surfactant; wherein the silicone-based biophotonic
membrane comprises between 80-99 wt % silicone phase and between
1-20 wt % thermogellable surfactant phase; wherein the
silicone-based biophotonic membrane when exposed to actinic light
having a wavelength of between about 400 nm and about 800 nm emits
fluorescence; wherein the silicone-based biophotonic membrane has a
light transmittance of between about 90% and about 100%: and
wherein the silicone-based biophotonic membrane has a thickness of
between about 0.5 mm and about 20 mm, a tensile strength of at
least about 50 kPa, and a tear strength of from about 0.1 N/mm to
about 5 N/mm.
2. The silicone-based biophotonic membrane of claim 1, wherein the
thermogellable surfactant phase is emulsified in the silicone
phase.
3. The silicone-based biophotonic membrane of claim 1, wherein the
surfactant comprises a block copolymer, wherein the block copolymer
comprises at least one hydrophobic block and at least one
hydrophilic block.
4. The silicone-based biophotonic membrane of claim 1, wherein the
surfactant comprises at least one sequence selected from the group
consisting of polyethylene glycol-propylene glycol ((PEG)-(PPG)),
polyethylene glycol-polylactic acid (PEG)-(PLA), polyethylene
glycol-poly(lactic-c-glycolic acid) (PEG)-(PLGA), and polyethyelene
glycol-polycaprolactone (PEG)-(PCL).
5. The silicone-based biophotonic membrane of claim 4, wherein the
surfactant is a poloxamer.
6. The silicone-based biophotonic membrane of claim 1, wherein the
thermogellable surfactant phase further comprises a surfactant
selected from the group consisting of cetyl trimethylammonium
bromide (CTAB) and sodium dodecyl sulfate (SDS).
7. The silicone-based biophotonic membrane of claim 1, wherein the
chromophore is a cationic chromophore selected from the group
consisting of cyanine, acridine and pyronine Y.
8. The silicone-based biophotonic membrane of claim 1, wherein the
chromophore is Eosin Y.
9. The silicone-based biophotonic membrane of claim 1, wherein the
thermogellable surfactant phase further comprises a stabilizer
selected from the group consisting of gelatin, hydroxylethyl
cellulose (HEC), carboxymethyl cellulose (CMC), and a thickening
agent.
10. The silicone-based biophotonic membrane of claim 1, wherein the
silicone phase comprises a polydimethylsiloxane polymer (PDMS).
11. The silicone-based biophotonic membrane of claim 10, wherein
the content of the PDMS in the silicone phase is from about 5 wt %
to about 100 wt %.
12. A method for biophotonic skin treatment, comprising: placing a
silicone-based biophotonic membrane over a target skin tissue,
wherein the silicone-based biophotonic membrane comprises a
silicone phase and a thermogellable surfactant phase, and wherein
the thermogellable surfactant phase comprises at least one
chromophore solubilized in a surfactant; wherein the silicone-based
biophotonic membrane comprises between 80-99 wt % silicone phase
and between 1-20 wt % thermogellable surfactant phase; and wherein
the silicone-based biophotonic membrane has a light transmittance
of between about 90% and about 100%; and wherein the silicone-based
biophotonic membrane has a thickness of between about 0.5 mm and
about 20 mm, a tensile strength of at least about 50 kPa, and a
tear strength of from about 0.1 N/mm to about 5 N/mm; and
illuminating said silicone-based biophotonic membrane with light
having a wavelength of between about 400 nm and about 800 nm;
wherein the silicone-based biophotonic membrane when exposed to the
actinic light having the wavelength that overlaps with the
absorption spectrum of the at least one chromophore emits
fluorescence.
13. The method of claim 12, wherein the skin treatment comprises
treating a skin disorder selected from the group consisting of
acne, eczema, psoriasis, and dermatitis.
14. The method of claim 12, wherein the skin treatment comprises
promoting skin rejuvenation.
15. The method of claim 12, wherein the silicone-based biophotonic
membrane is illuminated until the chromophore is at least partially
photobleached.
16. A method for promoting wound healing comprising: placing a
silicone-based biophotonic membrane over a wound, wherein the
silicone-based biophotonic membrane comprises a silicone phase and
a thermogellable surfactant phase, and wherein the thermogellable
surfactant phase comprises at least one chromophore solubilized in
a surfactant; wherein the silicone-based biophotonic membrane
comprises between 80-99 wt % silicone phase and between 1-20 wt %
thermogellable surfactant phase; wherein the silicone-based
biophotonic membrane has a light transmittance of between about 90%
and about 100%; and wherein the silicone-based biophotonic membrane
has a thickness of between about 0.5 mm and about 20 mm, a tensile
strength of at least about 50 kPa, and a tear strength of from
about 0.1 N/mm to about 5 N/mm; and illuminating said
silicone-based biophotonic membrane with light having a wavelength
of between about 400 nm and about 800 nm; wherein the
silicone-based biophotonic membrane when exposed to the actinic
light emits fluorescence.
17. The method of claim 16, wherein the method comprises treating
or preventing scarring.
Description
BACKGROUND OF THE DISCLOSURE
Phototherapy has recently been recognized as having wide range of
applications in both the medical and cosmetic fields including use
in surgery, therapy and diagnostics. For example, phototherapy has
been used to treat cancers and tumors with lessened invasiveness,
to disinfect target sites as an antimicrobial treatment, to promote
wound healing, and for facial skin rejuvenation.
Photodynamic therapy is a type of phototherapy involving the
application of a photosensitive agent to target tissue then
exposing the target tissue to a light source after a determined
period of time during which the photosensitizer is absorbed by the
target tissue. Such regimens, however, are often associated with
undesired side-effects, including systemic or localized toxicity to
the patient or damage to non-targeted tissue. Moreover, such
existing regimens often demonstrate low therapeutic efficacy due
to, for example, the poor selectivity of the photosensitive agents
into the target tissues.
Silicones are compounds based on alkylsiloxane or organosiloxane
and include polydimethylenesiloxane (PDMS), that have been
recognized as biocompatible and have been successfully used in
medical applications over the last six decades (Curtis et al., In
Biomaterials Science 2.sup.nd Edition, 2004). PDMS-based
compositions are widely used in personal care and skin topical
applications because they are non-irritating, non-sensitizing, and
meet the strict standards imposed by the US and European regulatory
agencies.
Therefore, it is an object of the present disclosure to provide new
and improved silicone based-compositions useful in phototherapy and
methods for their use.
SUMMARY OF THE DISCLOSURE
The present disclosure provides silicone-based biophotonic
compositions and methods useful in phototherapy. In particular, the
biophotonic compositions of the present disclosure include a
silicone matrix, and at least one chromophore, wherein the at least
one chromophore can absorb and emit light from within the
biophotonic composition, and which may be useful for cosmetic or
medical treatment of a human or animal tissue.
In one aspect, there is provided a silicone-based biophotonic
composition comprising a silicone phase and a surfactant phase,
wherein the surfactant phase comprises at least one chromophore
solubilized in a surfactant. In some embodiments, the surfactant
phase is emulsified in the silicone phase. In certain embodiments,
the silicone phase is a continuous phase. In some embodiments, the
surfactant is a block copolymer. The block copolymer may comprise
at least one hydrophobic block and at least one hydrophilic block.
In some embodiments the surfactant is thermogellable.
In certain embodiments of any of the foregoing or following, the
surfactant comprises at least one sequence of polyethylene
glycol-polypropylene glycol ((PEG)-(PPG)). In a further embodiment
the surfactant is a triblock copolymer or poloxomer of the formula
(PEG)-(PPG)-(PEG). In yet another embodiment, the surfactant is
Pluronic F127.
In certain embodiments of any of the foregoing or following, the
surfactant comprises at least one sequence of polyethylene
glycol-polylactic acid ((PEG)-(PLA)). In some embodiments the
surfactant comprises at least one sequence of polyethyelene
glycol-poly(lactic-c -glycolic acid) ((PEG)-(PLGA)). In some
embodiments the surfactant comprises at least one sequence of
polyethyelene glycol-polycaprolactone ((PEG)-(PCL)). In a further
embodiment the surfactant is a triblock copolymer or poloxomer of
the formula A-B-A or B-A-B, wherein A is PEG and B is PLA or PLGA
or PCL.
In certain embodiments of any of the foregoing or following, the
silicone phase comprises silicone. In certain embodiments, the
silicone may be a silicone elastomer. In certain embodiments, the
silicone comprises a polydimethylsiloxane. In certain embodiments,
the silicone comprises Sylgard.RTM. 184. In certain embodiments the
silicone comprises a mixture of Sylgard.RTM. 184 and Sylgard.RTM.
527. In a further embodiment the silicone comprises a mixture of
about 15% Sylgard.RTM. 184 and about 85% Sylgard.RTM. 527. In
certain embodiments, the mixture of Sylgard.RTM. 184 and
Sylgard.RTM. 527 provides for a silicone-based biophotonic
composition in a membrane form having an elasticity and a tackiness
which may be well suited to skin applications. Specifically, the
elasticity may allow for a greater ease of manipulation of the
silicone-based biophotonic membrane, and the tackiness (stickiness)
may allow for the membrane to stay where it is placed during a
treatment procedure as may be provided for in the present
disclosure.
In certain embodiments of any of the foregoing or following, the
silicone-based biophotonic composition comprises 80 wt % silicone
phase and about 20 wt % surfactant phase. In some embodiments the
silicone-based biophotonic compsotion comprises a silicone
phase/surfactant phase wt % composition of about 60/40 wt %, or
about 65/55 wt %, or about 70/30 wt %, or about 75/25 wt %, or
about 80/20 wt %, or about 85/15 wt % or about 90/10 wt %.
In certain embodiments of any of the foregoing or following, the at
least one chromophore is water soluble and is solubilized in the
surfactant phase. The at least one chromophore may be a
fluorophore. In certain embodiments, the chromophore can absorb
and/or emit light. In some embodiments, the light absorbed and/or
emitted by the chromophore is in the visible range of the
electromagnetic spectrum. In some embodiments, the light absorbed
and/or emitted by the chromophore is in the range of about 400 nm
to about 750 nm. In certain embodiments, the chromophore can emit
light from around 500 nm to about 700 nm. In some embodiments, the
chromophore or the fluorophore is a xanthene dye. The xanthene dye
may be selected from Eosin Y, Eosin B, Erythrosine B, Fluorescein,
Rose Bengal and Phloxin B.
In certain embodiments of any of the foregoing or following, the
surfactant phase of the silicone-based biophotonic composition
further comprises a stabilizer. In further embodiments the
stabilizer comprises gelatin, hydroxyethyl cellulose ether (HEC),
carboxymethyl cellulose (CMC) or any other thickening agent.
In certain embodiments of any of the foregoing or following, the
silicone-based biophotonic composition is at least substantially
translucent. The silicone-based biophotonic composition may be
transparent. In some embodiments, the silicone-based biophotonic
composition has a translucency of at least about 40%, about 50%,
about 60%, about 70%, or about 80% in a visible range. Preferably,
the light transmission through the composition is measured in the
absence of the at least one chromophore.
In certain embodiments, the composition is in the form of a
membrane. In other embodiments, the composition is in the form of a
spreadable gel.
In certain embodiments of any of the foregoing or following, the
surfactant phase further comprises an oxidizing agent. The
oxidizing agent may comprise a peroxide, such as hydrogen peroxide,
urea peroxide and benzoyl peroxide, or any other oxidizing agent
which can modulate the light absorption and/or emission properties
of the at least one chromophore or which can oxidize or degrade the
chromophore. For example, in certain embodiments where a single use
of the composition is desired, a peroxide may be included in the
surfactant phase to ensure degradation of the at least one
chromophore within a single treatment time.
In certain embodiments of any of the foregoing or following, the
silicone-based biophotonic composition, for example in the form of
a silicone-based biophotonic membrane, has a thickness of about 0.1
mm to about 50 mm, about 0.5 mm to about 20 mm, or about 1 mm to
about 10 mm, or about 1 mm to about 5 mm. In some embodiments, the
biophotonic composition is in the form of a gel that is applied at
a thickness of about 0.1 mm to about 50 mm, about 0.5 mm to about
20 mm, or about 1 mm to about 10 mm, or about 1 mm to about 5
mm.
In certain embodiments of any of the foregoing or following, the
silicone-based composition, for example in the form of a
silicone-based biophotonic membrane, has a removeable cover for
covering one or both sides of the membrane. The removeable cover
may be peelable. The removeable cover may comprise a sheet or a
film of material, such as paper or foil. In certain embodiments,
the removeable cover is opaque and can protect the membrane from
illumination until the treatment time. The cover may be partially
removeable. In certain embodiments, the cover may be re-applicable
to the membrane surface, such as after a treatment time, in order
to protect the membrane from further illumination in between
treatments.
In certain embodiments of any of the foregoing or following, the
surfactant phase is homogenously distributed within the silicone
phase and is nano and/or micro-sized. It can be considered as
micro-emulsified. The surfactant phase is not visibly detectable by
eye. In other words, the membrane appears by eye as one phase.
The silicone-based biophotonic composition of any aspects or
embodiments of the disclosure may be used for cosmetic or medical
treatment of tissue. In some embodiments, the cosmetic treatment is
skin rejuvenation and conditioning, and the medical treatment is
wound healing, periodontal treatment or acne treatment or treatment
of other skin conditions including eczema, psoriasis or dermatitis.
In some aspects, the silicone-based biophotonic membrane is used
for modulating inflammation, for modulating collagen synthesis, or
for promoting angiogenesis.
The present disclosure also provides methods for biophotonic
treatment comprising applying the silicone-based biophotonic
composition of the disclosure to a target tissue and illuminating
the composition with light.
From one aspect, there is provided a method for biophotonic
treatment of a skin disorder wherein the method comprises placing a
silicone-based biophotonic composition of the disclosure on or over
a target skin tissue, and illuminating said silicone-based
biophotonic composition with light having a wavelength that
overlaps with an absorption spectrum of the at least one
chromophore. In some embodiments, the biophotonic composition emits
fluorescence at a wavelength and intensity that promotes healing of
said skin disorder. The skin disorder may be selected from eczema,
psoriasis or dermatitis.
From another aspect, there is provided a method for biophotonic
treatment of acne comprising: placing a silicone-based biophotonic
composition of the disclosure on or over a target skin tissue; and
illuminating said composition with light having a wavelength that
overlaps with an absorption spectrum of the at least one
chromophore. In some embodiments, the biophotonic composition emits
fluorescence at a wavelength and intensity that treats the
acne.
From another aspect, there is provided a method for promoting wound
healing comprising: placing a silicone-based biophotonic
composition of the disclosure on or over a wound and illuminating
said silicone-based biophotonic composition with light having a
wavelength that overlaps with an absorption spectrum of the at
least one chromophore. In some embodiments, the biophotonic
composition emits fluorescence at a wavelength and intensity that
promotes wound healing.
From another aspect, there is provided a method for biophotonic
tissue repair comprising: placing a silicone-based biophotonic
composition of the disclosure on or over a target tissue; and
illuminating said silicone-based biophotonic composition with light
having a wavelength that overlaps with an absorption spectrum of
the at least one chromophore. In some embodiments, the biophotonic
composition emits fluorescence at a wavelength and intensity that
promotes tissue repair.
From another aspect, there is provided a method for promoting skin
rejuvenation comprising: placing a silicone-based biophotonic
composition of the disclosure on or over a target skin tissue; and
illuminating said silicone-based biophotonic composition with light
having a wavelength that overlaps with an absorption spectrum of
the at least one chromophore. In some embodiments, the biophotonic
composition emits fluorescence at a wavelength and intensity that
promotes skin rejuvenation. Promoting skin rejuvenation may
comprise promoting collagen synthesis.
From another aspect, there is provided a method for preventing or
treating scarring comprising: placing a silicone-based biophotonic
composition of the disclosure on or over a tissue scar; and
illuminating said silicone-based biophotonic composition with light
having a wavelength that overlaps with an absorption spectrum of
the at least one chromophore. In some embodiments, silicone-based
biophotonic composition emits fluorescence at a wavelength and
intensity that diminishes or prevents scarring.
In certain embodiments, the silicone-based biophotonic composition
is left in place after illumination. In certain embodiments, the
silicone-based biophotonic composition is re-illuminated. In some
embodiments, the chromophore at least partially photobleaches
during or after illumination. In certain embodiments, the
silicone-based biophotonic composition is illuminated until the
chromophore is at least partially photobleached.
In certain embodiments of any of the foregoing or following, the
light has a peak wavelength between about 400 nm and about 750 nm.
The light may have a peak wavelength between about 400 nm and about
500 nm.
In certain embodiments of any of the foregoing or following, the
light is from a direct light source such as a lamp. The lamp may be
an LED lamp. In certain embodiments, the light is from an ambient
light source.
In certain embodiments of any of the foregoing or following, said
silicone-based biophotonic composition is illuminated by a direct
light source for about 1 minute to greater than 75 minutes, about 1
minute to about 75 minutes, about 1 minute to about 60 minutes,
about 1 minute to about 55 minutes, about 1 minute to about 50
minutes, about 1 minute to about 45 minutes, about 1 minute to
about 40 minutes, about 1 minute to about 35 minutes, about 1
minute to about 30 minutes, about 1 minute to about 25 minutes,
about 1 minute to about 20 minutes, about 1 minute to about 15
minutes, about 1 minute to about 10 minutes, or about 1 minute to
about 5 minutes.
From a further aspect, there is provided use of the compositions
described above for tissue repair; for wound healing; for
preventing or treating scars; for skin rejuvenation; for treating
skin conditions such as acne, eczema, psoriasis or dermatitis; for
modulating inflammation; or for modulating collagen synthesis.
BRIEF DESCRIPTION OF THE DRAWINGS
Further aspects and advantages of the present invention will become
better understood with reference to the description in association
with the following in which:
FIG. 1 illustrates the light emission spectra of one embodiment of
the present disclosure comprising a silicone-based biophotonic
composition in a form of a membrane during 0-5 minutes of
illumination.
FIG. 2 illustrates the light emission spectra of the membrane of
FIG. 1 during 5-10 minutes of illumination.
FIG. 3 illustrates the light emission spectra of the membrane of
FIG. 1 during 10-15 minutes of illumination.
FIG. 4, panels A and B illustrate photobleaching of the membrane of
FIG. 1 over the indicated time period.
FIG. 5 illustrates a graph indicating a reduced dermal thickness of
scars in a dermal fibrotic mouse-human skin graft model after
treatment with a silicone-based biophotonic composition of the
present description.
FIG. 6 illustrates a graph indicating an improved collagen
remodeling, as measured with a collagen orientation index (COI), in
a dermal fibrotic mouse--human skin graft model after treatment
with a silicone-based biophotonic composition of the present
description.
DETAILED DESCRIPTION
(1) Overview
The present disclosure provides silicone-based biophotonic
compositions and uses thereof. Biophotonic therapy using these
compositions would combine the beneficial effects of topical
silicone compositions with the photobiostimulation induced by the
fluorescent light generated by the chromophore(s) upon illumination
of the compositions. Furthermore, in certain embodiments,
phototherapy using the silicone-based biophotonic membranes of the
present disclosure will for instance rejuvenate the skin by, e.g.,
promoting collagen synthesis, promote wound healing, prevent or
treat scars or to treat a skin conditions such as acne, eczema,
psoriasis, and treat periodontitis.
(2) Definitions
Before continuing to describe the present disclosure in further
detail, it is to be understood that this disclosure is not limited
to specific compositions or process steps, as such may vary. It
must be noted that, as used in this specification and the appended
claims, the singular form "a", "an" and "the" include plural
referents unless the context clearly dictates otherwise.
As used herein, the term "about" in the context of a given value or
range refers to a value or range that is within 20%, preferably
within 10%, and more preferably within 5% of the given value or
range.
It is convenient to point out here that "and/or" where used herein
is to be taken as specific disclosure of each of the two specified
features or components with or without the other. For example "A
and/or B" is to be taken as specific disclosure of each of (i) A,
(ii) B and (iii) A and B, just as if each is set out individually
herein.
"Biophotonic" means the generation, manipulation, detection and
application of photons in a biologically relevant context. In other
words, biophotonic compositions exert their physiological effects
primarily due to the generation and manipulation of photons.
"Topical application" or "topical uses" means application to body
surfaces, such as the skin, mucous membranes, vagina, oral cavity,
internal surgical wound sites, and the like.
"Emulsion" shall be understood as referring to a temporary or
permanent dispersion of one liquid phase within a second liquid
phase. Generally one of the phases is an aqueous solution, and the
other a water-immiscible liquid. The water-immiscible liquid is
generally referred to as the continuous phase. In this disclosure,
the continuous phase comprises a silicone and is referred to as a
silicone phase. Moreover, in this disclosure, the aqueous phase
comprises a surfactant and is referred to as a surfactant
phase.
Terms "chromophore" and "photoactivator" are used herein
interchangeably. A chromophore means a chemical compound, when
contacted by light irradiation, is capable of absorbing the light.
The chromophore readily undergoes photoexcitation and can transfer
its energy to other molecules or emit it as light
(fluorescence).
"Photobleaching" or "photobleaches" means the photochemical
destruction of a chromophore. A chromophore may fully or partially
photobleach.
The term "actinic light" is intended to mean light energy emitted
from a specific light source (e.g. lamp, LED, or laser) and capable
of being absorbed by matter (e.g. the chromophore or
photoactivator). Terms "actinic light" and "light" are used herein
interchangeably. In a preferred embodiment, the actinic light is
visible light.
"Skin rejuvenation" means a process of reducing, diminishing,
retarding or reversing one or more signs of skin aging or generally
improving the condition of skin. For instance, skin rejuvenation
may include increasing luminosity of the skin, reducing pore size,
reducing fine lines or wrinkles, improving thin and transparent
skin, improving firmness, improving sagging skin (such as that
produced by bone loss), improving dry skin (which might itch),
reducing or reversing freckles, reducing or preventing the
appearance of age spots, spider veins, rough and leathery skin,
fine wrinkles that disappear when stretched, reducing loose skin,
or improving a blotchy complexion. According to the present
disclosure, one or more of the above conditions may be improved or
one or more signs of aging may be reduced, diminished, retarded or
even reversed by certain embodiments of the compositions, methods
and uses of the present disclosure.
"Wound" means an injury to any tissue, including for example,
acute, subacute, delayed or difficult to heal wounds, and chronic
wounds. Examples of wounds may include both open and closed wounds.
Wounds include, for example, amputations, burns, incisions,
excisions, lesions, lacerations, abrasions, puncture or penetrating
wounds, surgical wounds, amputations, contusions, hematomas,
crushing injuries, ulcers (such as for example pressure, diabetic,
venous or arterial), scarring, and wounds caused by periodontitis
(inflammation of the periodontium).
Features and advantages of the subject matter hereof will become
more apparent in light of the following detailed description of
selected embodiments, as illustrated in the accompanying figures.
As will be realized, the subject matter disclosed and claimed is
capable of modifications in various respects, all without departing
from the scope of the claims. Accordingly, the drawings and the
description are to be regarded as illustrative in nature, and not
as restrictive and the full scope of the subject matter is set
forth in the claims.
(3) Silicone-Based Biophotonic Compositions
The present disclosure provides, in a broad sense, silicone-based
biophotonic compositions and methods of using silicone-based
biophotonic compositions. Silicone-based biophotonic compositions
can be, in a broad sense, activated by light (e.g., photons) of
specific wavelength. A silicone-based biophotonic composition
according to various embodiments of the present disclosure
comprises a silicone phase and a surfactant phase, with at least
one chromophore solubilized in the surfactant phase. In some
embodiments, the surfactant phase is emulsified in the silicone
phase. The chromophore in the silicone-based biophotonic
composition may be activated by light. This activation accelerates
the dispersion of light energy, leading to light carrying on a
therapeutic effect on its own, and/or to the photochemical
activation of other agents contained in the composition (e.g.,
acceleration in the breakdown process of peroxide (an oxidant or
oxidizing agent) when such compound is present in the composition
or in contact with the composition, leading to the formation of
oxygen radicals, such as singlet oxygen). This may lead to the
breakdown of the chromophore and, in some embodiments, ensure that
the silicone-based biophotonic composition, for example in the form
of a membrane, is for single-use.
When a chromophore absorbs a photon of a certain wavelength, it
becomes excited. This is an unstable condition and the molecule
tries to return to the ground state, giving away the excess energy.
For some chromophores, it is favorable to emit the excess energy as
light when returning to the ground state. This process is called
fluorescence. The peak wavelength of the emitted fluorescence is
shifted towards longer wavelengths compared to the absorption
wavelengths due to loss of energy in the conversion process. This
is called the Stokes' shift. In the proper environment (e.g., in a
biophotonic composition) much of this energy is transferred to the
other components of the biophotonic composition or to the treatment
site directly.
Without being bound to theory, it is thought that fluorescent light
emitted by photoactivated chromophores may have therapeutic
properties due to its femto-, pico-, or nano-second emission
properties which may be recognized by biological cells and tissues,
leading to favourable biomodulation. Furthermore, generally, the
emitted fluorescent light has a longer wavelength and hence a
deeper penetration into the tissue than the activating light.
Irradiating tissue with such a broad range of wavelength, including
in some embodiments the activating light which passes through the
composition, may have different and complementary effects on the
cells and tissues. In other words, chromophores are used in the
silicone-based biophotonic compositions of the present disclosure
for therapeutic effect on tissues. This is a distinct application
of these photoactive agents and differs from the use of
chromophores as simple stains or as catalysts for
photo-polymerization.
The silicone-based biophotonic compositions of the present
disclosure may have topical uses such as a mask or a wound
dressing. In some embodiments, the silicone-based biophotonic
compositions are cohesive. The cohesive nature of these
silicone-based biophotonic compositions may provide ease of removal
from the site of treatment and hence provide for a convenient ease
of use. Additionally or alternatively, the silicone-based
biophotonic compositions of the present disclosure have functional
and structural properties and these properties may also be used to
define and describe the compositions. Individual components of the
silicone-based biophotonic compositions of the present disclosure,
including chromophores, surfactants, silicone, and other optional
ingredients, are detailed below.
(a) Chromophores
Suitable chromophores can be fluorescent compounds (or stains)
(also known as "fluorochromes" or "fluorophores"). Other dye groups
or dyes (biological and histological dyes, food colorings,
carotenoids, and other dyes) can also be used. Suitable
photoactivators can be those that are Generally Regarded As Safe
(GRAS). Advantageously, photoactivators which are not well
tolerated by the skin or other tissues can be included in the
biophotonic composition of the present disclosure, as in certain
embodiments, the photoactivators are encapsulated within the
surfactant phase of the emulsion in the silicone continuous
phase.
In certain embodiments, the chromophore is one which undergoes
partial or complete photobleaching upon application of light. In
some embodiments, the chromophore absorbs at a wavelength in the
range of the visible spectrum, such as at a wavelength of about
380-800 nm, 380-700, 400-800, or 380-600 nm. In other embodiments,
the chromophore absorbs at a wavelength of about 200-800 nm,
200-700 nm, 200-600 nm or 200-500 nm. In one embodiment, the
chromophore absorbs at a wavelength of about 200-600 nm. In some
embodiments, the chromophore absorbs light at a wavelength of about
200-300 nm, 250-350 nm, 300-400 nm, 350-450 nm, 400-500 nm, 450-650
nm, 600-700 nm, 650-750 nm or 700-800 nm.
It will be appreciated to those skilled in the art that optical
properties of a particular chromophore may vary depending on the
chromophore's surrounding medium. Therefore, as used herein, a
particular chromophore's absorption and/or emission wavelength (or
spectrum) corresponds to the wavelengths (or spectrum) measured in
a biophotonic composition of the present disclosure.
The silicone-based biophotonic composition disclosed herein may
include at least one additional chromophore or second chromophore.
Combining chromophores may increase photo-absorption by the
combined dye molecules and enhance absorption and
photo-biomodulation selectivity. This creates multiple
possibilities of generating new photosensitive, and/or selective
chromophores mixtures. Thus, in certain embodiments, silicone-based
biophotonic compositions of the disclosure include more than one
chromophore, and when illuminated with light, energy transfer can
occur between the chromophores. This process, known as resonance
energy transfer, is a widely prevalent photophysical process
through which an excited `donor` chromophore (also referred to
herein as first chromophore) transfers its excitation energy to an
`acceptor` chromophore (also referred to herein as second
chromophore). The efficiency and directedness of resonance energy
transfer depends on the spectral features of donor and acceptor
chromophores. In particular, the flow of energy between
chromophores is dependent on a spectral overlap reflecting the
relative positioning and shapes of the absorption and emission
spectra. More specifically, for energy transfer to occur, the
emission spectrum of the donor chromophore must overlap with the
absorption spectrum of the acceptor chromophore.
Energy transfer manifests itself through decrease or quenching of
the donor emission and a reduction of excited state lifetime
accompanied also by an increase in acceptor emission intensity. To
enhance the energy transfer efficiency, the donor chromophore
should have good abilities to absorb photons and emit photons.
Furthermore, the more overlap there is between the donor
chromophore's emission spectra and the acceptor chromophore's
absorption spectra, the better a donor chromophore can transfer
energy to the acceptor chromophore.
Accordingly, in embodiments comprising a mixture of chromophores,
the first chromophore has an emission spectrum that overlaps at
least about 80%, 50%, 40%, 30%, 20% or 10% with an absorption
spectrum of the second chromophore. In one embodiment, the first
chromophore has an emission spectrum that overlaps at least about
20% with an absorption spectrum of the second chromophore. In some
embodiments, the first chromophore has an emission spectrum that
overlaps at least 1-10%, 5-15%, 10-20%, 15-25%, 20-30%, 25-35%,
30-40%, 35-45%, 50-60%, 55-65%, 60-70% or 70-80% with an absorption
spectrum of the second chromophore.
% spectral overlap, as used herein, means the % overlap of a donor
chromophore's emission wavelength range with an acceptor
chromophore's absorption wavelength rage, measured at spectral full
width quarter maximum (FWQM). In some embodiments, the second
chromophore absorbs at a wavelength in the range of the visible
spectrum. In certain embodiments, the second chromophore has an
absorption wavelength that is relatively longer than that of the
first chromophore within the range of about 50-250, 25-150 or
10-100 nm.
The chromophore may be present in an amount of about 0.001-40% per
weight of the composition or of the surfactant phase. In certain
embodiments, the at least one chromophore is present in an amount
of about 0.001-3%, 0.001-0.01%, 0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%,
2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%,
20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or
35-40% per weight of the silicone-based biophotonic composition or
of the surfactant phase.
When present, the second chromophore may be present in an amount of
about 0.001-40% per weight of the silicone-based biophotonic
composition or of the surfactant phase. In certain embodiments, the
second chromophore is present in an amount of about 0.001-3%,
0.001-0.01%, 0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%, 2.5-7.5%, 5-10%,
7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%,
22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40% per
weight of the silicone-based biophotonic composition or of the
surfactant phase. In certain embodiments, the total weight per
weight of chromophore or combination of chromophores may be in the
amount of about 0.005-1%, 0.05-2%, 1-5%, 2.5-7.5%, 5-10%,
7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%,
22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40.001%
per weight of the silicone-based biophotonic composition of the
surfactant phase.
The concentration of the chromophore to be used can be selected
based on the desired intensity and duration of the biophotonic
activity from the silicone-based biophotonic composition, and on
the desired medical or cosmetic effect. For example, some dyes such
as xanthene dyes reach a `saturation concentration` after which
further increases in concentration do not provide substantially
higher emitted fluorescence. Further increasing the chromophore
concentration above the saturation concentration can reduce the
amount of activating light passing through the matrix. Therefore,
if more fluorescence is required for a certain application than
activating light, a high concentration of chromophore can be used.
However, if a balance is required between the emitted fluorescence
and the activating light, a concentration close to or lower than
the saturation concentration can be chosen.
Suitable chromophores that may be used in the silicone-based
biophotonic compositions of the present disclosure include, but are
not limited to the following:
Chlorophyll Dyes
Exemplary chlorophyll dyes include but are not limited to
chlorophyll a; chlorophyll b; chlorophyllin; bacteriochlorophyll a;
bacteriochlorophyll b; bacteriochlorophyll c; bacteriochlorophyll
d; protochlorophyll; protochlorophyll a; amphiphilic chlorophyll
derivative 1; and amphiphilic chlorophyll derivative 2.
Xanthene Derivatives Exemplary xanthene dyes include, but are not
limited to, esosin B, eosin B
(4',5'-dibromo,2',7'-dinitr-o-fluorescein, dianion); Eosin Y; eosin
Y (2',4',5',7'-tetrabromo-fluorescein, dianion); eosin
(2',4',5',7'-tetrabromo-fluorescein, dianion); eosin
(2',4',5',7'-tetrabromo-fluorescein, dianion) methyl ester; eosin
(2',4',5',7'-tetrabromo-fluorescein, monoanion) p-isopropylbenzyl
ester; eosin derivative (2',7'-dibromo-fluorescein, dianion); eosin
derivative (4',5'-dibromo-fluorescein, dianion); eosin derivative
(2',7'-dichloro-fluorescein, dianion); eosin derivative
(4',5'-dichloro-fluorescein, dianion); eosin derivative
(2',7'-diiodo-fluorescein, dianion); eosin derivative
(4',5'-diiodo-fluorescein, dianion); eosin derivative
(tribromo-fluorescein, dianion); eosin derivative
(2',4',5',7'-tetrachlor-o-fluorescein, dianion); eosin; eosin
dicetylpyridinium chloride ion pair; erythrosin B
(2',4',5',7'-tetraiodo-fluorescein, dianion); erythrosin;
erythrosin dianion; erythiosin B; fluorescein; fluorescein dianion;
phloxin B (2',4',5',7'-tetrabromo-3,4,5,6-tetrachloro-fluorescein,
dianion); phloxin B (tetrachloro-tetrabromo-fluorescein); phloxine
B; rose bengal
(3,4,5,6-tetrachloro-2',4',5',7'-tetraiodofluorescein, dianion);
pyronin G, pyronin J, pyronin Y; Rhodamine dyes such as rhodamines
include 4,5-dibromo-rhodamine methyl ester; 4,5-dibromo-rhodamine
n-butyl ester; rhodamine 101 methyl ester; rhodamine 123; rhodamine
6G; rhodamine 6G hexyl ester; tetrabromo-rhodamine 123; and
tetramethyl-rhodamine ethyl ester.
Methylene Blue Dyes
Exemplary methylene blue derivatives include but are not limited to
1-methyl methylene blue; 1,9-dimethyl methylene blue; methylene
blue; methylene violet; bromomethylene violet; 4-iodomethylene
violet;
1,9-dimethyl-3-dimethyl-amino-7-diethyl-amino-phenothiazine; and
1,9-dimethyl-3-diethylamino-7-dibutyl-amino-phenot-hiazine.
Azo Dyes
Exemplary azo (or diazo-) dyes include but are not limited to
methyl violet, neutral red, para red (pigment red 1), amaranth
(Azorubine S), Carmoisine (azorubine, food red 3, acid red 14),
allura red AC (FD&C 40), tartrazine (FD&C Yellow 5), orange
G (acid orange 10), Ponceau 4R (food red 7), methyl red (acid red
2), and murexide-ammonium purpurate.
In some aspects of the disclosure, the one or more chromophores of
the silicone-based biophotonic compositions disclosed herein can be
independently selected from any of Acid black 1, Acid blue 22, Acid
blue 93, Acid fuchsin, Acid green, Acid green 1, Acid green 5, Acid
magenta, Acid orange 10, Acid red 26, Acid red 29, Acid red 44,
Acid red 51, Acid red 66, Acid red 87, Acid red 91, Acid red 92,
Acid red 94, Acid red 101, Acid red 103, Acid roseine, Acid rubin,
Acid violet 19, Acid yellow 1, Acid yellow 9, Acid yellow 23, Acid
yellow 24, Acid yellow 36, Acid yellow 73, Acid yellow S, Acridine
orange, Acriflavine, Alcian blue, Alcian yellow, Alcohol soluble
eosin, Alizarin, Alizarin blue 2RC, Alizarin carmine, Alizarin
cyanin BBS, Alizarol cyanin R, Alizarin red S, Alizarin purpurin,
Aluminon, Amido black 10B, Amidoschwarz, Aniline blue WS,
Anthracene blue SWR, Auramine O, Azocannine B, Azocarmine G, Azoic
diazo 5, Azoic diazo 48, Azure A, Azure B, Azure C, Basic blue 8,
Basic blue 9, Basic blue 12, Basic blue 15, Basic blue 17, Basic
blue 20, Basic blue 26, Basic brown 1, Basic fuchsin, Basic green
4, Basic orange 14, Basic red 2, Basic red 5, Basic red 9, Basic
violet 2, Basic violet 3, Basic violet 4, Basic violet 10, Basic
violet 14, Basic yellow 1, Basic yellow 2, Biebrich scarlet,
Bismarck brown Y, Brilliant crystal scarlet 6R, Calcium red,
Carmine, Carminic acid, Celestine blue B, China blue, Cochineal,
Coelestine blue, Chrome violet CG, Chromotrope 2R, Chromoxane
cyanin R, Congo corinth, Congo red, Cotton blue, Cotton red,
Croceine scarlet, Crocin, Crystal ponceau 6R, Crystal violet,
Dahlia, Diamond green B, Direct blue 14, Direct blue 58, Direct
red, Direct red 10, Direct red 28, Direct red 80, Direct yellow 7,
Eosin B, Eosin Bluish, Eosin, Eosin Y, Eosin yellowish, Eosinol,
Erie garnet B, Eriochrome cyanin R, Erythrosin B, Ethyl eosin,
Ethyl green, Ethyl violet, Evans blue, Fast blue B, Fast green FCF,
Fast red B, Fast yellow, Fluorescein, Food green 3, Gallein,
Gallamine blue, Gallocyanin, Gentian violet, Haematein, Haematine,
Haematoxylin, Helio fast rubin BBL, Helvetia blue, Hematein,
Hematine, Hematoxylin, Hoffman's violet, Imperial red, Indocyanin
Green, Ingrain blue, Ingrain blue 1, Ingrain yellow 1, INT, Kermes,
Kermesic acid, Kernechtrot, Lac, Laccaic acid, Lauth's violet,
Light green, Lissamine green SF, Luxol fast blue, Magenta O,
Magenta I, Magenta II, Magenta III, Malachite green, Manchester
brown, Martius yellow, Merbromin, Mercurochrome, Metanil yellow,
Methylene azure A, Methylene azure B, Methylene azure C, Methylene
blue, Methyl blue, Methyl green, Methyl violet, Methyl violet 2B,
Methyl violet 10B, Mordant blue 3, Mordant blue 10, Mordant blue
14, Mordant blue 23, Mordant blue 32, Mordant blue 45, Mordant red
3, Mordant red 11, Mordant violet 25, Mordant violet 39 Naphthol
blue black, Naphthol green B, Naphthol yellow S, Natural black 1,
Natural green 3(chlorophyllin), Natural red, Natural red 3, Natural
red 4, Natural red 8, Natural red 16, Natural red 25, Natural red
28, Natural yellow 6, NBT, Neutral red, New fuchsin, Niagara blue
3B, Night blue, Nitro BT, Nitro blue tetrazolium, Nuclear fast red,
Orange G, Orcein, Pararosanilin, Phloxine B, Picric acid, Ponceau
2R, Ponceau 6R, Ponceau B, Ponceau de Xylidine, Ponceau S, Primula,
Purpurin, Pyronin B, phycobilins, Phycocyanins, Phycoerythrins.
Phycoerythrincyanin (PEC), Phthalocyanines, Pyronin G, Pyronin Y,
Quinine, Rhodamine B, Rosanilin, Rose bengal, Saffron, Safranin O,
Scarlet R, Scarlet red, Scharlach R, Shellac, Sirius red F3B,
Solochrome cyanin R, Soluble blue, Spirit soluble eosin, Sulfur
yellow S, Swiss blue, Tartrazine, Thioflavine S, Thioflavine T,
Thionin, Toluidine blue, Toluyline red, Tropaeolin G, Trypaflavine,
Trypan blue, Uranin, Victoria blue 4R, Victoria blue B, Victoria
green B, Vitamin B, Water blue I, Water soluble eosin, Xylidine
ponceau, or Yellowish eosin.
In certain embodiments, the silicone-based biophotonic compositions
of the present disclosure includes any of the chromophores listed
above, or a combination thereof, so as to provide a synergistic
biophotonic effect at the application site.
Without being bound to any particular theory, a synergistic effect
of the chromophore combinations means that the biophotonic effect
is greater than the sum of their individual effects.
Advantageously, this may translate to increased reactivity of the
biophotonic composition, faster or improved treatment time. Also,
the treatment conditions need not be altered to achieve the same or
better treatment results, such as time of exposure to light, power
of light source used, and wavelength of light used. In other words,
use of synergistic combinations of chromophores may allow the same
or better treatment without necessitating a longer time of exposure
to a light source, a higher power light source or a light source
with different wavelengths.
In some embodiments, the composition includes Eosin Y as a first
chromophore and any one or more of Rose Bengal, Fluorescein,
Erythrosine, Phloxine B, chlorophyll as a second chromophore. It is
believed that these combinations have a synergistic effect as they
can transfer energy to one another when activated due in part to
overlaps or close proximity of their absorption and emission
spectra. This transferred energy is then emitted as fluorescence
and/or leads to production of reactive oxygen species. This
absorbed and re-emitted light is thought to be transmitted
throughout the composition, and also to be transmitted into the
site of treatment.
In further embodiments, the silicone-based biophotonic composition
may include, for example, the following synergistic combinations:
Eosin Y and Fluorescein; Fluorescein and Rose Bengal; Erythrosine
in combination with Eosin Y, Rose Bengal or Fluorescein; Phloxine B
in combination with one or more of Eosin Y, Rose Bengal,
Fluorescein and Erythrosine.
By means of synergistic effects of the chromophore combinations in
the silicone-based biophotonic composition, chromophores which
cannot normally be activated by an activating light (such as a blue
light from an LED), can be activated through energy transfer from
chromophores which are activated by the activating light. In this
way, the different properties of photoactivated chromophores can be
harnessed and tailored according to the cosmetic or the medical
therapy required.
For example, Rose Bengal can generate a high yield of singlet
oxygen when activated in the presence of molecular oxygen, however
it has a low quantum yield in terms of emitted fluorescent light.
Rose Bengal has a peak absorption around 540 nm and so can be
activated by green light. Eosin Y has a high quantum yield and can
be activated by blue light. By combining Rose Bengal with Eosin Y,
one obtains a composition which can emit therapeutic fluorescent
light and generate singlet oxygen when activated by blue light. In
this case, the blue light photoactivates Eosin Y, which transfers
some of its energy to Rose Bengal as well as emitting some energy
as fluorescence.
In some embodiments, the chromophore or chromophores are selected
such that their emitted fluorescent light, on photoactivation, is
within one or more of the green, yellow, orange, red and infrared
portions of the electromagnetic spectrum, for example having a peak
wavelength within the range of about 490 nm to about 800 nm. In
certain embodiments, the emitted fluorescent light has a power
density of between 0.005 to about 10 mW/cm.sup.2, about 0.5 to
about 5 mW/cm.sup.2.
(b) Surfactant Phase
The silicone-based biophotonic compositions of the present
disclosure comprise a surfactant phase. The surfactant may be
present in an amount of at least 5%, 10%, 15%, 20%, 25%, or 30% of
the total composition. In certain embodiments, the surfactant phase
comprises a block copolymer. The term "block copolymer" as used
herein refers to a copolymer comprised of 2 or more blocks (or
segments) of different homopolymers. The term homopolymer refers to
a polymer comprised of a single monomer. Many variations of block
copolymers are possible including simple diblock polymers with an
A-B architecture and triblock polymers with A-B-A, B-A-B or A-B-C
architectures and more complicated block copolymers are known. In
addition, unless otherwise indicated herein, the repetition number
and type of the monomers or repeating units constituting the block
copolymer are not particularly limited. For example, when one
denotes the monomeric repeating units as "a" and "b", it is meant
herein that this copolymer includes not only a random copolymer
having the average composition of (a).sub.m(b).sub.n, but also a
diblock copolymer of the composition (a).sub.m(b).sub.n, and a
triblock copolymer of the composition (a).sub.l(b).sub.m(a).sub.n,
or the like. In the formulae above, l, m, and n represent the
number of repeating units and are positive numbers.
In certain embodiments of any of the foregoing or following the
block copolymer is biocompatible. A polymer is "biocompatible" in
that the polymer and degradation products thereof are substantially
non-toxic to cells or organisms, including non-carcinogenic and
non-immunogenic, and are cleared or otherwise degraded in a
biological system, such as an organism (patient) without
substantial toxic effect.
In certain embodiments the block copolymer of the surfactant phase
is from a group of tri-block copolymers designated Poloxamers.
Poloxamers are A-B-A block copolymers in which the A segment is a
hydrophilic polyethylene glycol (PEG) homopolymer and the B segment
is hydrophobic polypropylene glycol (PPG) homopolymer. PEG is also
known as polyethylene oxide (PEO) or polyoxyethylene (POE),
depending on its molecular weight. Additionally, PPG is also known
as polypropylene oxide (PPO), depending on its molecular weight.
Poloxamers are commercially available from BASF Corporation.
Poloxamers produce reverse thermal gelatin compositions, i.e., with
the characteristic that their viscosity increases with increasing
temperature up to a point from which viscosity again decreases.
Depending on the relative size of the blocks the copolymer can be a
solid, liquid or paste. In certain embodiments of the disclosure,
the poloxamer is Pluronic.RTM. F127 (also known as Poloxamer 407).
In some embodiments of the silicone-based biophotonic composition
may comprise Pluronic.RTM. F127 in the amount of 1-40 wt % of the
total composition. In some embodiments of the silicone-based
biophotonic composition may comprise 1-5 wt %, 2.5-7.5 wt %, 5-10
wt %, 7.5-12.5 wt %, 10-15 wt %, 12.5-17.5 wt %, 15-20 wt %, 20-25
wt %, 25-30 wt %, 30-35 wt %, 35-40 wt % pluronic. In certain
embodiments Pluronic.RTM. F127 is present in the amount of 2-8 wt %
of the total composition of the silicone-based biophotonic
composition.
In certain embodiments of the disclosure the surfactant phase
comprises a block copolymer comprising at least an A-B unit,
wherein A is PEG and B is polylactic acid (PLA), or polyglycolic
acid (PGA) or poly(lactic-co-glycolic acid) (PLGA) or
polycaprolactone (PCL) or polydioxanone (PDO).
Since the PEG blocks contribute hydrophilicity to the polymer,
increasing the length of the PEG blocks or the total amount of PEG
in the polymer will tend to make the polymer more hydrophilic.
Depending on the amounts and proportions of the other components of
the polymer, the desired overall hydrophilicity, and the nature and
chemical functional groups of any chromophore that may be included
in a formulation of the polymer, a skilled person can readily
adjust the length (or MW) of the PEG blocks used and/or the total
amount of PEG incorporated into the polymer, in order to obtain a
polymer having the desired physical and chemical
characteristics.
The total amount of PEG in the polymer may be about 80 wt % or
less, 75 wt % or less, 70 wt % or less, 65 wt % or less, about 60
wt % or less, about 55 wt % or less, or about 50 wt % or less. In
particular embodiments, the total amount of PEG is about 55 wt %,
56 wt %, 57 wt %, 58 wt %, 59 wt %, 60 wt %, 61 wt %, 62 wt %, 63
wt %, 64 wt %, 65 wt %, 66 wt %, 67 wt %, 68 wt %, 69 wt %, or
about 70 wt %. Unless otherwise specified, a weight percentage of a
particular component of the polymer means that the total weight of
the polymer is made up of the specified percentage of monomers of
that component. For example, 65 wt % PEG means that 65% of the
weight of the polymer is made up of PEG monomers, which monomers
are linked into blocks of varying lengths, which blocks are
distributed along the length of polymer, including in a random
distribution.
The total amount of PPG or PLA or PLGA or PCL or PDO present in the
block copolymer may be about 50 wt % or less, about 45 wt % or
less, about 40 wt % or less, about 35 wt % or less, about 30 wt %
or less, about 25 wt % or less, or about 20 wt % or less.
The surfactant phase may also include thickening agents or
stabilizers such as gelatin and/or modified celluloses such as
hydroxyethyl cellulose (HEC) and carboxymethyl cellulose (CMD),
and/or polysaccharides such as xanthan gum, guar gum, and/or
starches and/or any other thickening agent. In certain embodiments
of the disclosure, the stabilizer or thickening agent may comprise
gelatin. For example, the surfactant phase may comprise about 0-5
wt %, about 5-25 wt %, about 0-15 wt %, or about 10-20 wt %
gelatin.
Surfactants and/or stabilizers may be selected according to effects
they will have on the optical transparency of the biophotonic
membrane. The silicone-based biophotonic composition should be able
to transmit sufficient light to activate the at least one
chromophore and, in embodiments where fluorescence is emitted by
the activated chromophore, the surfactant phase should also be able
to transmit the emitted fluorescent light to tissues.
(c) Silicone Phase
The silicone-based biophotonic compositions of the present
disclosure comprise a continuous phase of silicone. Silicones are
synthetic polymers containing chains consisting of (--Si--O--)
repeating unit with two organic groups attached directly to the Si
atom. In certain embodiments, the silicone is a
polydimethylsiloxane (PDMS) fluid (Me.sub.2SiO).sub.n or a
PDMS-based gel or PDMS-based elastomer.
Non-limiting examples of PDMS polymers include those sold under the
trademark Sylgard, and particularly Sylgard 182, Sylgard 184,
Sylgard 186 and Sylgard 527. In certain embodiments, the silicone
phase of the silicone-based biophotonic composition can be prepared
by using commercial kits such as Sylgard.RTM. 184 Silicone
Elastomer kit. The kit consists in two-part liquid components, the
base (part A) and the curing agent or catalyst (part B), both based
on polydimethylsiloxane. When mixed at a ratio of 10(A)/1(B), the
mixture cures to a flexible and transparent elastomer.
Sylgard 184 is a silicone elastomer comprising a polydimethyl
siloxane and an organically-modified silica. Sylgard 184 is
prepared by combining a base (Part A) with a curing agent (Part B).
The base contains about >60 wt % dimethylvinyl-terminated
dimethyl siloxane, about 30 to 60 wt % dimethylvinylated and
trimethylated silica and about 1 to 5 wt % tetra(trimethylsiloxy)
silane. The curing agent contains about 40 to 70 wt % dimethyl,
methylhydrogen siloxane, about 15 to 40 wt %
dimethylvinyl-terminated dimethyl siloxane, about 10 to 30 wt %
dimethylvinylated and trimethylated silica and about 1 to 5 wt %
tetramethyl tetravinyl cyclotetrasiloxane.
In another embodiment, the silicone phase of the silicone-based
biophotonic composition can be prepared by using the Sylgard.RTM.
527 Silicone Gel kit, which allows the preparation of a soft and
sticky gel, when the two parts A and B are mixed at the ratio
1(A)/1(B). Parts A and B of Sylgard contain about 85 to 100 wt %
dimethylvinyl-terminated dimethyl siloxane and about 1 to 5 wt %
dimethyl, methylhydrogen siloxane.
In other embodiments, the silicone-based biophotonic composition
may be prepared in a manner to provide for tunable flexibility were
desired, for example a silicone-based biophotonic membrane having
tunable flexibility. One means of generating a tunable
silicone-based biophotonic membrane of the present disclosure is by
combining different ratios of commercially available PDMS such as
Sylgard.RTM. 184 and Sylgard.RTM. 527. In some embodiments the
silicone phase comprises Sylgard.RTM. 184 in the amount of 5-100 wt
% of the silicone phase. In certain embodiments of the present
disclosure the Sylgard.RTM. 184 is present in an amount of about
5-10 wt %, 10-15 wt %, 15-20 wt %, 20-25 wt %, 25-30 wt %, 30-35 wt
%, 35-40 wt %, 40-45 wt %, 45-50 wt %, 50-55 wt %, 55-60 wt %,
60-65 wt % 65-70 wt %, 70-75 wt %, 75-80 wt %, 80-85 wt %, 85-90 wt
%, 90-95 wt % or 95-100 wt % of the silicone phase. In certain
embodiments of the present disclosure, the silicone phase comprises
Sylgard.RTM. 527. In certain other embodiments of the present
disclosure, the Sylgard.RTM. 527 is present in an amount of about
5-10 wt %, 10-15 wt %, 15-20 wt %, 20-25 wt %, 25-30 wt %, 30-35 wt
%, 35-40 wt %, 40-45 wt %, 45-50 wt %, 50-55 wt %, 55-60 wt %,
60-65 wt % 65-70 wt %, 70-75 wt %, 75-80 wt %, 80-85 wt %, 85-90 wt
%, 90-95 wt % or 95-100 wt % of the silicone phase.
In one embodiment, the silicone phase of the silicone-based
biophotonic composition is a mixture using 15% Sylgard 184 and 85%
Sylgard 527.
(d) Oxidizing Agents and Antimicrobials
According to certain embodiments, the silicone-based biophotonic
composition of the present disclosure, or the surfactant phase of
these silicone-based biophotonic compositions, may optionally
comprise one or more additional components, such as oxygen-rich
compounds as a source of oxygen radicals. The oxygen-rich compounds
include but are not limited to peroxides, such as hydrogen
peroxide, benzoyl peroxide and urea peroxide. Peroxide compounds
are oxidants that contain the peroxy group (R--O--O--R), which is a
chainlike structure containing two oxygen atoms, each of which is
bonded to the other and a radical or some element. When a
silicone-based biophotonic composition of the present disclosure is
illuminated with light, the chromophores are excited to a higher
energy state. When the chromophores' electrons return to a lower
energy state, they emit photons with a lower energy level, thus
causing the emission of light of a longer wavelength (Stokes'
shift). Some of this energy may be transferred to the oxidizing
agent and may cause the formation of oxygen radicals, such as
singlet oxygen. These oxygen radicals may contribute to the
degradation of the chromophore.
Hydrogen peroxide (H.sub.2O.sub.2) is a powerful oxidizing agent,
and breaks down into water and oxygen and does not form any
persistent, toxic residual compound. A suitable range of
concentration over which hydrogen peroxide can be used in the
silicone-based biophotonic composition is from about 0.1% to about
3%, about 0.1 to 1.5%, about 0.1% to about 1%, about 1%, less than
about 1%.
Urea hydrogen peroxide (also known as urea peroxide, carbamide
peroxide or percarbamide) is soluble in water and contains
approximately 35% hydrogen peroxide. A suitable range of
concentration over which urea peroxide can be used in the
silicone-based biophotonic composition of the present disclosure is
less than about 0.25%, or less than about 0.3%, from 0.001 to
0.25%, or from about 0.3% to about 5%. Urea peroxide breaks down to
urea and hydrogen peroxide in a slow-release fashion that can be
accelerated with heat or photochemical reactions.
Benzoyl peroxide consists of two benzoyl groups (benzoic acid with
the H of the carboxylic acid removed) joined by a peroxide group. A
suitable range of concentration over which benzoyl peroxide can be
used in the silicone-based biophotonic composition is from about
2.5% to about 5%.
Antimicrobials kill microbes or inhibit their growth or
accumulation, and may optionally be included with the
silicone-based biophotonic compositions of the present disclosure.
Suitable antimicrobials for use in the methods and compositions of
the present disclosure include, but not limited to, hydrogen
peroxide, urea hydrogen peroxide, benzoyl peroxide, phenolic and
chlorinated phenolic and chlorinated phenolic compounds, resorcinol
and its derivatives, bisphenolic compounds, benzoic esters
(parabens), halogenated carbonilides, polymeric antimicrobial
agents, thazolines, trichloromethylthioimides, natural
antimicrobial agents (also referred to as "natural essential
oils"), metal salts, and broad-spectrum antibiotics.
Specific phenolic and chlorinated phenolic antimicrobial agents
that can be used in the disclosure include, but are not limited to:
phenol; 2-methyl phenol; 3-methyl phenol; 4-methyl phenol; 4-ethyl
phenol; 2,4-dimethyl phenol; 2,5-dimethyl phenol; 3,4-dimethyl
phenol; 2,6-dimethyl phenol; 4-n-propyl phenol; 4-n-butyl phenol;
4-n-amyl phenol; 4-tert-amyl phenol; 4-n-hexyl phenol; 4-n-heptyl
phenol; mono- and poly-alkyl and aromatic halophenols;
p-chlorophenyl; methyl p-chlorophenol; ethyl p-chlorophenol;
n-propyl p-chlorophenol; n-butyl p-chlorophenol; n-amyl
p-chlorophenol; sec-amyl p-chlorophenol; n-hexyl p-chlorophenol;
cyclohexyl p-chlorophenol; n-heptyl p-chlorophenol; n-octyl;
p-chlorophenol; o-chlorophenol; methyl o-chlorophenol; ethyl
o-chlorophenol; n-propyl o-chlorophenol; n-butyl o-chlorophenol;
n-amyl o-chlorophenol; tert-amyl o-chlorophenol; n-hexyl
o-chlorophenol; n-heptyl o-chlorophenol; o-benzyl p-chlorophenol;
o-benxyl-m-methyl p-chlorophenol; o-benzyl-m,m-dimethyl
p-chlorophenol; o-phenylethyl p-chlorophenol;
o-phenylethyl-m-methyl p-chlorophenol; 3-methyl p-chlorophenol
3,5-dimethyl p-chlorophenol, 6-ethyl-3-methyl p-chlorophenol,
6-n-propyl-3-methyl p-chlorophenol; 6-iso-propyl-3-methyl
p-chlorophenol; 2-ethyl-3,5-dimethyl p-chlorophenol;
6-sec-butyl-3-methyl p-chlorophenol; 2-iso-propyl-3,5-dimethyl
p-chlorophenol; 6-diethylmethyl-3-methyl p-chlorophenol;
6-iso-propyl-2-ethyl-3-methyl p-chlorophenol;
2-sec-amyl-3,5-dimethyl p-chlorophenol;
2-diethylmethyl-3,5-dimethyl p-chlorophenol; 6-sec-octyl-3-methyl
p-chlorophenol; p-chloro-m-cresol p-bromophenol; methyl
p-bromophenol; ethyl p-bromophenol; n-propyl p-bromophenol; n-butyl
p-bromophenol; n-amyl p-bromophenol; sec-amyl p-bromophenol;
n-hexyl p-bromophenol; cyclohexyl p-bromophenol; o-bromophenol;
tert-amyl o-bromophenol; n-hexyl o-bromophenol;
n-propyl-m,m-dimethyl o-bromophenol; 2-phenyl phenol;
4-chloro-2-methyl phenol; 4-chloro-3-methyl phenol;
4-chloro-3,5-dimethyl phenol; 2,4-dichloro-3,5-dimethylphenol;
3,4,5,6-tetabromo-2-methylphenol-; 5-methyl-2-pentylphenol;
4-isopropyl-3-methylphenol; para-chloro-metaxylenol (PCMX);
chlorothymol; phenoxyethanol; phenoxyisopropanol; and
5-chloro-2-hydroxydiphenylmethane.
Resorcinol and its derivatives can also be used as antimicrobial
agents. Specific resorcinol derivatives include, but are not
limited to: methyl resorcinol; ethyl resorcinol; n-propyl
resorcinol; n-butyl resorcinol; n-amyl resorcinol; n-hexyl
resorcinol; n-heptyl resorcinol; n-octyl resorcinol; n-nonyl
resorcinol; phenyl resorcinol; benzyl resorcinol; phenylethyl
resorcinol; phenylpropyl resorcinol; p-chlorobenzyl resorcinol;
5-chloro-2,4-dihydroxydiphenyl methane;
4'-chloro-2,4-dihydroxydiphenyl methane;
5-bromo-2,4-dihydroxydiphenyl methane; and
4'-bromo-2,4-dihydroxydiphenyl methane.
Specific bisphenolic antimicrobial agents that can be used in the
disclosure include, but are not limited to: 2,2'-methylene
bis-(4-chlorophenol); 2,4,4'trichloro-2'-hydroxy-diphenyl ether,
which is sold by Ciba Geigy, Florham Park, N.J. under the tradename
Triclosan.RTM.; 2,2'-methylene bis-(3,4,6-trichlorophenol);
2,2'-methylene bis-(4-chloro-6-bromophenol);
bis-(2-hydroxy-3,5-dichlorop-henyl) sulphide; and
bis-(2-hydroxy-5-chlorobenzyl)sulphide.
Specific benzoie esters (parabens) that can be used in the
disclosure include, but are not limited to: methylparaben;
propylparaben; butylparaben; ethylparaben; isopropylparaben;
isobutylparaben; benzylparaben; sodium methylparaben; and sodium
propylparaben.
Specific halogenated carbanilides that can be used in the
disclosure include, but are not limited to:
3,4,4'-trichlorocarbanilides, such as
3-(4-chlorophenyl)-1-(3,4-dichlorphenyl)urea sold under the
tradename Triclocarban.RTM. by Ciba-Geigy, Florham Park, N.J.;
3-trifluoromethyl-4,4'-dichlorocarbanilide; and
3,3',4-trichlorocarbanilide Specific polymeric antimicrobial agents
that can be used in the disclosure include, but are not limited to:
polyhexamethylene biguanide hydrochloride; and
poly(iminoimidocarbonyl iminoimidocarbonyl iminohexamethylene
hydrochloride), which is sold under the tradename Vantocil.RTM.
IB.
Specific thazolines that can be used in the disclosure include, but
are not limited to that sold under the tradename Micro-Check.RTM.
and 2-n-octyl-4-isothiazolin-3-one, which is sold under the
tradename Vinyzene.RTM. IT-3000 DIDP.
Specific trichloromethylthioimides that can be used in the
disclosure include, but are not limited to:
N-(trichloromethylthio)phthalimide, which is sold under the
tradename Fungitrol.RTM.; and
N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide, which is
sold under the tradename Vancide.RTM..
Specific natural antimicrobial agents that can be used in the
disclosure include, but are not limited to, oils of: anise; lemon;
orange; rosemary; wintergreen; thyme; lavender; cloves; hops; tea
tree; citronella; wheat; barley; lemongrass; cedar leaf; cedarwood;
cinnamon; fleagrass; geranium; sandalwood; violet; cranberry;
eucalyptus; vervain; peppermint; gum benzoin; basil; fennel; fir;
balsam; menthol; ocmea origanuin; hydastis; carradensis;
Berberidaceac daceae; Ratanhiae longa; and Curcuma longa. Also
included in this class of natural antimicrobial agents are the key
chemical components of the plant oils which have been found to
provide antimicrobial benefit. These chemicals include, but are not
limited to: anethol; catechole; camphene; thymol; eugenol;
eucalyptol; ferulic acid; farnesol; hinokitiol; tropolone;
limonene; menthol; methyl salicylate; carvacol; terpineol;
verbenone; berberine; ratanhiae extract; caryophellene oxide;
citronellic acid; curcumin; nerolidol; and geraniol.
Specific metal salts that can be used in the disclosure include,
but are not limited to, salts of metals in groups 3a-5a, 3b-7b, and
8 of the periodic table. Specific examples of metal salts include,
but are not limited to, salts of: aluminum; zirconium; zinc;
silver; gold; copper; lanthanum; tin; mercury; bismuth; selenium;
strontium; scandium; yttrium; cerium; praseodymiun; neodymium;
promethum; samarium; europium; gadolinium; terbium; dysprosium;
holmium; erbium; thalium; ytterbium; lutetium; and mixtures
thereof. An example of the metal-ion based antimicrobial agent is
sold under the tradename HealthShield.RTM., and is manufactured by
HealthShield Technology, Wakefield, Mass.
Specific broad-spectrum antimicrobial agents that can be used in
the disclosure include, but are not limited to, those that are
recited in other categories of antimicrobial agents herein.
Additional antimicrobial agents that can be used in the methods of
the disclosure include, but are not limited to: pyrithiones, and in
particular pyrithione-including zinc complexes such as that sold
under the tradename Octopirox.RTM.; dimethyidimethylol hydantoin,
which is sold under the tradename Glydant.RTM.;
methylchloroisothiazolinone/methylisothiazolinone, which is sold
under the tradename Kathon CG.RTM.; sodium sulfite; sodium
bisulfite; imidazolidinyl urea, which is sold under the tradename
Germall 115.RTM.; diazolidinyl urea, which is sold under the
tradename Germall 110; benzyl alcohol
v2-bromo-2-nitropropane-1,3-diol, which is sold under the tradename
Bronopol.RTM.; formalin or formaldehyde; iodopropenyl
butylcarbamate, which is sold under the tradename Polyphase
P100.RTM.; chloroacetamide; methanamine; methyldibromonitrile
glutaronitrile (1,2-dibromo-2,4-dicyanobutane), which is sold under
the tradename Tektamer.RTM.; glutaraldehyde;
5-bromo-5-nitro-1,3-dioxane, which is sold under the tradename
Bronidox.RTM.; phenethyl alcohol; o-phenylphenol/sodium
o-phenylphenol sodium hydroxymethylglycinate, which is sold under
the tradename Suttocide A.RTM.; polymethoxy bicyclic oxazolidine;
which is sold under the tradename Nuosept C.RTM.; dimethoxane;
thimersal; dichlorobenzyl alcohol; captan; chlorphenenesin;
dichlorophene; chlorbutanol; glyceryl laurate; halogenated diphenyl
ethers; 2,4,4'-trichloro-2'-hydroxy-diphenyl ether, which is sold
under the tradename Triclosan.RTM. and is available from
Ciba-Geigy, Florham Park, N.J.; and
2,2'-dihydroxy-5,5'-dibromo-diphenyl ether.
(4) Optical Properties of the Silicone-Based Biophotonic
Compositions
In certain embodiments, silicone-based biophotonic compositions of
the present disclosure are substantially transparent or
translucent. The % transmittance of the silicone-based biophotonic
composition can be measured in the range of wavelengths from 250 nm
to 800 nm using, for example, a Perkin-Elmer Lambda 9500 series
UV-visible spectrophotometer. In some embodiments, transmittance
within the visible range is measured and averaged. In some other
embodiments, transmittance of the silicone-based biophotonic
composition is measured with the chromophore omitted. As
transmittance is dependent upon thickness, the thickness of each
sample can be measured with calipers prior to loading in the
spectrophotometer. Transmittance values can be normalized according
to
.function..lamda..sigma..function..lamda..times..function..lamda.
##EQU00001##
where t.sub.1=actual specimen thickness, t.sub.2=thickness to which
transmittance measurements can be normalized. In the art,
transmittance measurements are usually normalized to 1 cm.
In some embodiments, the silicone-based biophotonic composition has
a transmittance that is more than about 20%, 30%, 40%, 50%, 60%,
70%, or 75% within the visible range. In some embodiments, the
transmittance exceeds 40%, 41%, 42%, 43%, 44%, or 45% within the
visible range. In some embodiments, the silicone-based biophotonic
composition has a light transmittance of about 40-100%, 45-100%,
50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%,
85-100%, 90-100%, or 95-100%.
(5) Forms of the Silicone-Based Biophotonic Compositions
The silicone-based biophotonic compositions of the present
disclosure may be in the form of a silicone-based biophotonic
membrane containing at least one chromophore.
The silicone-based biophotonic membranes of the present disclosure
may be deformable. They may be elastic or non-elastic (i.e.
flexible or rigid). The silicone-based biophotonic membrane, for
example, may be in a peel-off form (`peelable`) to provide ease and
speed of use. In certain embodiments, the tear strength and/or
tensile strength of the peel-off form is greater than its adhesion
strength. This may help handleability of the silicone-based
biophotonic membrane. It will be recognized by one of skill in the
art that the properties of the peel-off silicone-based biophotonic
membrane such as cohesiveness, flexibility, elasticity, tensile
strength, and tearing strength, can be determined and/or adjusted
by methods known in the art such as by selecting suitable
PDMS-based compositions and adapting their relative ratios.
The silicone-based biophotonic composition may be provided in a
pre-formed shape. In certain embodiments, the pre-formed shape is
in the form of, including, but not limited to, a film, a face mask,
a patch, a dressing, or bandage. In certain embodiments, the
pre-formed shapes can be customized for the individual user by
trimming to size. In certain embodiments, perforations are provided
around the perimeter of the pre-formed shape to facilitate
trimming. In certain embodiments, the pre-shaping can be performed
manually or by mechanical means such as 3-D printing. In the case
of the 3-D printing the size of the area to be treated can be
imaged, such as a wound or a face, then a 3-D printer configured to
build or form a cohesive silicone-based biophotonic composition to
match the size and shape of the imaged treatment area.
A silicone-based biophotonic composition of the disclosure can be
configured with a shape and/or size for application to a desired
portion of a subject's body. For example, the silicone-based
biophotonic composition can be shaped and sized to correspond with
a desired portion of the body to receive the biophotonic treatment.
Such a desired portion of skin can be selected from, but not
limited to, the group consisting of a skin, head, forehead, scalp,
nose, cheeks, lips, ears, face, neck, shoulder, arm pit, arm,
elbow, hand, finger, abdomen, chest, stomach, back, buttocks,
sacrum, genitals, legs, knee, feet, toes, nails, hair, any boney
prominences, and combinations thereof, and the like. Thus, the
silicone-based biophotonic composition of the disclosure can be
shaped and sized to be applied to any portion of skin on a
subject's body. For example, the silicone-based biophotonic
composition can be in the form of a sock, hat, glove or mitten
shaped form. In embodiments where the silicone-based biophotonic
composition is in a elastic, semi-rigid or rigid form, it may be
peeled-off without leaving any residue on the tissue.
In certain embodiments, the silicone-based biophotonic composition
is provided in the form of an elastic and peelable face mask, which
may be pre-formed. In other embodiments, the silicone-based
biophotonic composition is in the form of a non-elastic (rigid)
face mask, which may also be pre-formed. The mask can have openings
for one or more of the eyes, nose and mouth. In a further
embodiment, the openings are protected with a covering, or the
exposed skin such as on the nose, lips or eyes are protected using
for example cocoa butter. In certain embodiments, the pre-formed
face mask is provided in the form of multiple parts, e.g., an upper
face part and a lower face part. In certain embodiments, the uneven
proximity of the face to a light source is compensated for, e.g.,
by adjusting the thickness of the mask, or by adjusting the amount
of chromophore in the different areas of the mask, or by blocking
the skin in closest proximity to the light. In certain embodiments,
the pre-formed shapes come in a one-size fits all form.
In certain embodiments, the silicone-based biophotonic composition
is in the form of a wound dressing or a bandage. It may be used on
a wound to prevent or limit scar formation, or on an existing scar
to diminish the appearance of the scar.
In certain aspects, the mask (or patch) is not pre-formed and is
applied e.g., by spreading a silicone-based biophotonic composition
making up the mask (or patch), on the skin or target tissue, or
alternatively by smearing, dabbing or rolling the composition on
target tissue. It can then be converted to a peel-off form after
application, by means such as, but not limited to, drying or
inducing a change in temperature upon application to the skin or
tissue. After use, the mask (or patch) can then be peeled off
without leaving any flakes on the skin or tissue, preferably
without wiping or washing.
The silicone-based biophotonic compositions of the present
disclosure may, for example when provided in the form of a
silicone-based biophotonic membrane, mask or dressing, have a
thickness of from about 0.1 mm to about 50 mm, about 0.5 mm to
about 20 mm, or about 1 mm to about 10 mm. It will be appreciated
that the thickness will vary based on the intended use. In some
embodiments, the thickness ranges from about 0.1-1 mm. In some
embodiments, the thickness ranges from about 0.5-1.5 mm, about 1-2
mm, about 1.5-2.5 mm, about 2-3 mm, about 2.5-3.5 mm, about 3-4 mm,
about 3.5-4.5 mm, about 4-5 mm, about 4.5-5.5 mm, about 5-6 mm,
about 5.5-6.5 mm, about 6-7 mm, about 6.5-7.5 mm, about 7-8 mm,
about 7.5-8.5 mm, about 8-9 mm, about 8.5-9.5, about 9-10 mm, about
10-11 mm, about 11-12 mm, about 12-13 mm, about 13-14 mm, about
14-15 mm, about 15-16 mm, about 16-17 mm, about 17-18 mm, about
18-19 mm, about 19-20 mm, about 20-22 mm, about 22-24 mm, about
24-26 mm, about 26-28 mm, about 28-30 mm, about 30-35 mm, about
35-40 mm, about 40-45 mm, about 45-50 mm.
The tensile strength of the silicone-based biophotonic compositions
will vary based on the intended use. The tensile strength can be
determined by performing a tensile test and recording the force and
displacement. These are then converted to stress (using cross
sectional area) and strain; the highest point of the stress-strain
curve is the "ultimate tensile strength." In some embodiments, for
example when in the form of a silicone-based biophotonic membrane,
tensile strength can be characterized using a 500N capacity
tabletop mechanical testing system (#5942R4910, Instron.RTM.) with
a 5N maximum static load cell (#102608, Instron). Pneumatic side
action grips can be used to secure the samples (#2712-019,
Instron). In some embodiments, a constant extension rate (for
example, of about 2 mm/min) until failure can be applied and the
tensile strength is calculated from the stress vs. strain data
plots. In some embodiments, the tensile strength can be measured
using methods as described in or equivalent to those described in
American Society for Testing and Materials tensile testing methods
such as ASTM D638, ASTM D882 and ASTM D412.
In some embodiments, the silicone-based biophotonic composition has
a tensile strength that is at least about 50 kPa, at least about
100 kPa, at least about 200 kPa, at least about 300 kPa, at least
about 400 kPa, at least about 500 kPa, at least about 600 kPa, at
least about 700 kPa, at least about 800 kPa, at least about 900
kPa, at least about 1 MPa, at least about 2 MPa or at least about 3
MPa, or at least about 5 MPa, or at least about 6 MPa. In some
embodiments, the tensile strength of the silicone-based biophotonic
composition is up to about 10 MPa.
The tear strength of the silicone-based biophotonic composition
will vary depending on the intended use. The tear strength property
of the silicone-based biophotonic composition, for example when
provided in the form of a silicone-based biophotonic membrane, can
be tested using a 500N capacity tabletop mechanical testing system
(#5942R4910, Instron) with a 5N maximum static load cell (#102608,
Instron). Pneumatic side action grips can be used to secure the
samples (#2712-019, Instron). Samples can be tested with a constant
extension rate (for example, of about 2 mm/min) until failure. In
accordance with the invention, tear strength is calculated as the
force at failure divided by the average thickness (N/mm).
In some embodiments, the silicone-based biophotonic composition has
a tear strength of from about 0.1 N/mm to about 5 N/mm. In some
embodiments, the tear strength is from about 0.1 N/mm to about 0.5
N/mm, from about 0.25 N/mm to about 0.75 N/mm, from about 0.5 N/mm
to about 1.0 N/mm, from about 0.75 N/mm to about 1.25 N/mm, from
about 1.0 N/mm to about 1.5 N/mm, from about 1.5 N/mm to about 2.0
N/mm, from about 2.0 N/mm to about 2.5 N/mm, from about 2.5 N/mm to
about 3.0 N/mm, from about 3.0 N/mm to about 3.5 N/mm, from about
3.5 N/mm to about 4.0 N/mm, from about 4.0 N/mm to about 4.5 N/mm,
from about 4.5 N/mm to about 5.0 N/mm.
The adhesion strength of the silicone-based biophotonic composition
will vary depending on the intended use. Adhesion strength can be
determined in accordance with ASTM D-3330-78, PSTC-101 and is a
measure of the force required to remove a silicone-based
biophotonic composition from a test panel at a specific angle and
rate of removal. In some embodiments, a predetermined size of the
silicone-based biophotonic composition, for example a
silicone-based biophotonic membrane, is applied to a horizontal
surface of a clean glass test plate. A hard rubber roller is used
to firmly apply a piece of the silicone-based biophotonic membrane
and remove all discontinuities and entrapped air. The free end of
the piece of silicone-based biophotonic membrane is then doubled
back nearly touching itself so that the angle of removal of the
piece from the glass plate will be 180 degrees. The free end of the
piece of silicone-based biophotonic membrane is attached to the
adhesion tester scale (e.g. an Instron tensile tester or Harvey
tensile tester). The test plate is then clamped in the jaws of the
tensile testing machine capable of moving the plate away from the
scale at a predetermined constant rate. The scale reading in kg is
recorded as the silicone-based biophotonic membrane is peeled from
the glass surface.
In some embodiments, the adhesion strength can be measured by
taking into account the static friction of the silicone-based
biophotonic composition. For some embodiments of the silicone-based
biophotonic compositions of the present disclosure, the adhesive
properties are linked to their levels of static friction, or
stiction. In these cases, the adhesion strength can be measured by
placing a sample of the silicone-based biophotonic composition such
as a silicone-based biophotonic membrane on a test surface and
pulling one end of the sample at an angle of approximately
0.degree. (substantially parallel to the surface) whilst applying a
known downward force (e.g. a weight) on the sample and measuring
the weight at which the sample slips from the surface. The normal
force F.sub.n, is the force exerted by each surface on the other in
a perpendicular (normal) direction to the surface and is calculated
by multiplying the combined weight of the sample and the weight by
the gravity constant (g) (9.8 m/s.sup.2). The sample with the
weight on top is then pulled away from a balance until the sample
slips from the surface and the weight is recorded on the scale. The
weight recorded on the scale is equivalent to the force required to
overcome the friction. The force of friction (F.sub.f) is then
calculated by multiplying the weight recorded on the scale by g.
Since F.sub.f.ltoreq..mu.F.sub.n (Coulomb's friction law), the
friction coefficient .mu. can be obtained by dividing
F.sub.f/F.sub.n. The stress required to shear a material from a
surface (adhesion strength) can then be calculated from the
friction coefficient, .mu., by multiplying the weight of the
material by the friction coefficient.
In some embodiments, the silicone-based biophotonic composition has
an adhesion strength that is less than its tensile strength or its
tear strength.
In some embodiments, the silicone-based biophotonic composition has
an adhesion strength of from about 0.01 N/mm to about 0.60 N/mm. In
some embodiments, the adhesion strength is from about 0.20 N/mm to
about 0.40 N/mm, or from about 0.25 N/mm to about 0.35 N/mm. In
some embodiments, the adhesion strength is less than about 0.10
N/mm, less than about 0.15 N/mm, less than about 0.20 N/mm, less
than about 0.25 N/mm, less than about 0.30 N/mm, less than about
0.35 N/mm, less than about 0.40 N/mm, less than about 0.45 N/mm,
less than about 0.55 N/mm or less than about 0.60 N/mm
(6) Methods of Use
The silicone-based biophotonic compositions of the present
disclosure may have cosmetic and/or medical benefits. They may be
used to promote skin rejuvenation and skin conditioning, or to
promote the treatment of a skin disorder such as acne, eczema,
dermatitis or psoriasis, or to promote tissue repair, modulate
inflammation, modulate collagen synthesis, reduce or avoid
scarring, or promote wound healing including reducing depth of
periodontitis pockets. In certain embodiments, the silicone-based
biophotonic composition of the disclosure maybe used to treat acute
inflammation, which may present itself as pain, heat, redness,
swelling and loss of function, and which includes those seen in
allergic reactions such as insect bites e.g.; mosquito, bees,
wasps, poison ivy, or post-ablative treatment.
Accordingly, in certain embodiments, the present disclosure
provides a method for treating acute inflammation.
In certain embodiments, the present disclosure provides a method
for providing skin rejuvenation or for improving skin condition,
treating a skin disorder, preventing or treating scarring, and/or
accelerating wound healing and/or tissue repair, the method
comprising: applying a silicone-based biophotonic composition of
the present disclosure to the area of the skin or tissue in need of
treatment, and illuminating the silicone-based biophotonic
composition with light having a wavelength that overlaps with an
absorption spectrum of the chromophore(s) present in the
composition.
In the methods of the present disclosure, any source of actinic
light can be used. Any type of halogen, LED or plasma arc lamp, or
laser may be suitable. The primary characteristic of suitable
sources of actinic light will be that they emit light in a
wavelength (or wavelengths) appropriate for activating the one or
more photoactivators present in the composition. In one embodiment,
an argon laser is used. In another embodiment, a potassium-titanyl
phosphate (KTP) laser (e.g. a GreenLight.TM. laser) is used. In yet
another embodiment, a LED lamp such as a photocuring device is the
source of the actinic light. In yet another embodiment, the source
of the actinic light is a source of light having a wavelength
between about 200 to 800 nm. In another embodiment, the source of
the actinic light is a source of visible light having a wavelength
between about 400 and 600 nm. In another embodiment, the source of
the actinic light is a source of visible light having a wavelength
between about 400 and 700 nm. In yet another embodiment, the source
of the actinic light is blue light. In yet another embodiment, the
source of the actinic light is red light. In yet another
embodiment, the source of the actinic light is green light.
Furthermore, the source of actinic light should have a suitable
power density. Suitable power density for non-collimated light
sources (LED, halogen or plasma lamps) are in the range from about
0.1 mW/cm.sup.2 to about 200 mW/cm.sup.2. Suitable power density
for laser light sources are in the range from about 0.5 mW/cm.sup.2
to about 0.8 mW/cm.sup.2.
In some embodiments of the methods of the present disclosure, the
light has an energy at the subject's skin surface of between about
0.1 mW/cm.sup.2 and about 500 mW/cm.sup.2, or 0.1-300 mW/cm.sup.2,
or 0.1-200 mW/cm.sup.2, wherein the energy applied depends at least
on the condition being treated, the wavelength of the light, the
distance of the skin from the light source and the thickness of the
biophotonic material. In certain embodiments, the light at the
subject's skin is between about 1-40 mW/cm.sup.2, or 20-60
mW/cm.sup.2, or 40-80 mW/cm.sup.2, or 60-100 mW/cm.sup.2, or 80-120
mW/cm.sup.2, or 100-140 mW/cm.sup.2, or 30-180 mW/cm.sup.2, or
120-160 mW/cm.sup.2, or 140-180 mW/cm.sup.2, or 160-200
mW/cm.sup.2, or 110-240 mW/cm.sup.2, or 110-150 mW/cm.sup.2, or
190-240 mW/cm.sup.2.
The activation of the chromophore(s) within the silicone-based
biophotonic composition may take place almost immediately on
illumination (femto- or pico seconds). A prolonged exposure period
may be beneficial to exploit the synergistic effects of the
absorbed, reflected and reemitted light of the silicone-based
biophotonic composition of the present disclosure and its
interaction with the tissue being treated. In one embodiment, the
time of exposure of the tissue or skin or silicone-based
biophotonic composition to actinic light is a period between 0.01
minutes and 90 minutes. In another embodiment, the time of exposure
of the tissue or skin or silicone-based biophotonic composition to
actinic light is a period between 1 minute and 5 minutes. In some
other embodiments, the silicone-based biophotonic composition is
illuminated for a period between 1 minute and 3 minutes. In certain
embodiments, light is applied for a period of 1-30 seconds, 15-45
seconds, 30-60 seconds, 0.75-1.5 minutes, 1-2 minutes, 1.5-2.5
minutes, 2-3 minutes, 2.5-3.5 minutes, 3-4 minutes, 3.5-4.5
minutes, 4-5 minutes, 5-10 minutes, 10-15 minutes, 15-20 minutes,
or 20-30 minutes. The treatment time may range up to about 90
minutes, about 80 minutes, about 70 minutes, about 60 minutes,
about 50 minutes, about 40 minutes or about 30 minutes. It will be
appreciated that the treatment time can be adjusted in order to
maintain a dosage by adjusting the rate of fluence delivered to a
treatment area. For example, the delivered fluence may be about 4
to about 60 J/cm.sup.2, about 10 to about 60 J/cm.sup.2, about 10
to about 50 J/cm.sup.2, about 10 to about 40 J/cm.sup.2, about 10
to about 30 J/cm.sup.2, about 20 to about 40 J/cm.sup.2, about 15
J/cm.sup.2 to 25 J/cm.sup.2, or about 10 to about 20
J/cm.sup.2.
In certain embodiments, the silicone-based biophotonic composition
may be re-illuminated at certain intervals. In yet another
embodiment, the source of actinic light is in continuous motion
over the treated area for the appropriate time of exposure. In yet
another embodiment, the silicone-based biophotonic composition may
be illuminated until the silicone-based biophotonic composition is
at least partially photobleached or fully photobleached.
In certain embodiments, the chromophore(s) may be photoexcited by
ambient light including from the sun and overhead lighting. In
certain embodiments, the chromophore(s) may be photoactivated by
light in the visible range of the electromagnetic spectrum. The
light may be emitted by any light source such as sunlight, light
bulb, an LED device, electronic display screens such as on a
television, computer, telephone, mobile device, flashlights on
mobile devices. In the methods of the present disclosure, any
source of light can be used. For example, a combination of ambient
light and direct sunlight or direct artificial light may be used.
Ambient light can include overhead lighting such as LED bulbs,
fluorescent bulbs etc, and indirect sunlight.
In the methods of the present disclosure, the silicone-based
biophotonic composition may be removed from the skin following
application of light. In other embodiments, the silicone-based
biophotonic composition is left on the tissue for an extended
period of time and re-activated with direct or ambient light at
appropriate times to treat the condition.
In certain embodiments of any of the foregoing or following, the
silicone-based biophotonic composition, such as a silicone-based
biophotonic membrane, has a removable cover for covering one or
both sides of the membrane. The removable cover may be peelable.
The removable cover may comprise a sheet or a film of material,
such as paper or foil. In certain embodiments, the removable cover
is opaque and can protect the membrane from illumination until the
treatment time. The cover may be partially removable. In certain
embodiments, the cover may be re-applicable to the membrane
surface, such as after a treatment time, in order to protect the
membrane from further illumination in between treatments.
In certain embodiments of the method of the present disclosure, the
silicone-based biophotonic composition may be applied to the
tissue, such as on the face, once, twice, three times, four times,
five times or six times a week, daily, or at any other frequency.
The total treatment time may be one week, two weeks, three weeks,
four weeks, five weeks, six weeks, seven weeks, eight weeks, nine
weeks, ten weeks, eleven weeks, twelve weeks, or any other length
of time deemed appropriate. In certain embodiments, the total
tissue area to be treated may be split into separate areas (cheeks,
forehead), and each area treated separately. For example, the
silicone-based biophotonic composition may be applied topically to
a first portion, and that portion illuminated with light, and the
composition then removed. Then the silicone-based biophotonic
composition is applied to a second portion, illuminated and
removed. Finally, the silicone-based biophotonic composition is
applied to a third portion, illuminated and removed.
In certain embodiments, the silicone-based biophotonic composition
can be used following wound closure to optimize scar revision. In
this case, the silicone-based biophotonic composition may be
applied at regular intervals such as once a week, or at an interval
deemed appropriate by the physician.
In certain embodiments, the silicone-based biophotonic composition
may be used following ablative skin rejuvenation treatment to
maintain the condition of the treated skin. In this case, the
silicone-based biophotonic composition may be applied at regular
intervals such as once a week, or at an interval deemed appropriate
by the physician.
In the methods of the present disclosure, additional components may
optionally be included with the silicone-based biophotonic
compositions or used in combination with the silicone-based
biophotonic compositions. Such additional components may include,
but are not limited to, healing factors, antimicrobials,
oxygen-rich agents, wrinkle fillers such as botox, hyaluronic acid
and polylactic acid, fungal, anti-bacterial, anti-viral agents
and/or agents that promote collagen synthesis. These additional
components may be applied to the skin in a topical fashion, prior
to, at the same time of, and/or after topical application of the
silicone-based biophotonic compositions of the present disclosure.
Suitable healing factors comprise compounds that promote or enhance
the healing or regenerative process of the tissues on the
application site. During the photoactivation of a silicone-based
biophotonic composition of the present disclosure, there may be an
increase of the absorption of molecules of such additional
components at the treatment site by the skin or the mucosa. Healing
factors may also modulate the biophotonic effect resulting from the
silicone-based biophotonic composition. Suitable healing factors
include, but are not limited to glucosamines, allantoins, saffron,
agents that promote collagen synthesis, anti-fungal,
anti-bacterial, anti-viral agents and wound healing factors such as
growth factors.
(i) Skin Rejuvenation
The silicone-based biophotonic compositions of the present
disclosure may be useful in promoting skin rejuvenation or
improving skin condition and appearance. The dermis is the second
layer of skin, containing the structural elements of the skin, the
connective tissue. There are various types of connective tissue
with different functions. Elastin fibers give the skin its
elasticity, and collagen gives the skin its strength.
The junction between the dermis and the epidermis is an important
structure. The dermal-epidermal junction interlocks forming
finger-like epidermal ridges. The cells of the epidermis receive
their nutrients from the blood vessels in the dermis. The epidermal
ridges increase the surface area of the epidermis that is exposed
to these blood vessels and the needed nutrients.
The aging of skin comes with significant physiological changes to
the skin. The generation of new skin cells slows down, and the
epidermal ridges of the dermal-epidermal junction flatten out.
While the number of elastin fibers increases, their structure and
coherence decreases. Also the amount of collagen and the thickness
of the dermis decrease with the ageing of the skin.
Collagen is a major component of the skin's extracellular matrix,
providing a structural framework. During the aging process, the
decrease of collagen synthesis and insolubilization of collagen
fibers contribute to a thinning of the dermis and loss of the
skin's biomechanical properties.
The physiological changes to the skin result in noticeable aging
symptoms often referred to as chronological-, intrinsic- and
photo-aging. The skin becomes drier, roughness and scaling
increase, the appearance becomes duller, and most obviously fine
lines and wrinkles appear. Other symptoms or signs of skin aging
include, but are not limited to, thinning and transparent skin,
loss of underlying fat (leading to hollowed cheeks and eye sockets
as well as noticeable loss of firmness on the hands and neck), bone
loss (such that bones shrink away from the skin due to bone loss,
which causes sagging skin), dry skin (which might itch), inability
to sweat sufficiently to cool the skin, unwanted facial hair,
freckles, age spots, spider veins, rough and leathery skin, fine
wrinkles that disappear when stretched, loose skin, a blotchy
complexion.
The dermal-epidermal junction is a basement membrane that separates
the keratinocytes in the epidermis from the extracellular matrix,
which lies below in the dermis. This membrane consists of two
layers: the basal lamina in contact with the keratinocytes, and the
underlying reticular lamina in contact with the extracellular
matrix. The basal lamina is rich in collagen type IV and laminin,
molecules that play a role in providing a structural network and
bioadhesive properties for cell attachment.
Laminin is a glycoprotein that only exists in basement membranes.
It is composed of three polypeptide chains (alpha, beta and gamma)
arranged in the shape of an asymmetric cross and held together by
disulfide bonds. The three chains exist as different subtypes which
result in twelve different isoforms for laminin, including
Laminin-1 and Laminin-5.
The dermis is anchored to hemidesmosomes, specific junction points
located on the keratinocytes, which consist of a-integrins and
other proteins, at the basal membrane keratinocytes by type VII
collagen fibrils. Laminins, and particularly Laminin-5, constitute
the real anchor point between hemidesmosomal transmembrane proteins
in basal keratinocytes and type VII collagen.
Laminin-5 synthesis and type VII collagen expression have been
proven to decrease in aged skin. This causes a loss of contact
between dermis and epidermis, and results in the skin losing
elasticity and becoming saggy.
Recently another type of wrinkles, generally referred to as
expression wrinkles, received general recognition. Expression
wrinkles result from a loss of resilience, particularly in the
dermis, because of which the skin is no longer able to resume its
original state when facial muscles which produce facial
expressions.
The silicone-based biophotonic compsoitions of the present
disclosure and methods of the present disclosure promote skin
rejuvenation. In certain embodiments, the silicone-based
biophotonic compositions and methods of the present disclosure may
promote skin conditioning such as skin luminosity, reduction of
pore size, reducing blotchiness, making even skin tone, reducing
dryness, and tightening of the skin. In certain embodiments, the
silicone-based biophotonic compositions and methods of the present
disclosure may promote collagen synthesis. In certain other
embodiments, the silicone-based biophotonic compositions and
methods of the present disclosure may reduce, diminish, retard or
even reverse one or more signs of skin aging including, but not
limited to, appearance of fine lines or wrinkles, thin and
transparent skin, loss of underlying fat (leading to hollowed
cheeks and eye sockets as well as noticeable loss of firmness on
the hands and neck), skin aging due bone loss (wherein bones shrink
away from the skin due to bone loss, which causes sagging skin),
dry skin (which might itch), inability to sweat sufficiently to
cool the skin, unwanted facial hair, freckles, age spots, spider
veins, rough and leathery skin, fine wrinkles that disappear when
stretched, loose skin, or a blotchy complexion. In certain
embodiments, the silicone-based biophotonic compositions and
methods of the present disclosure may induce a reduction in pore
size, enhance sculpturing of skin subsections, and/or enhance skin
translucence.
In certain embodiments, the silicone-based biophotonic composition
may be used in conjunction with collagen promoting agents. Agents
that promote collagen synthesis (i.e., pro-collagen synthesis
agents) include amino acids, peptides, proteins, lipids, small
chemical molecules, natural products and extracts from natural
products.
For instance, it has been discovered that intake of vitamin C,
iron, and collagen can effectively increase the amount of collagen
in skin or bone. See, e.g., U.S. Patent Application Publication
20090069217. Examples of the vitamin C include an ascorbic acid
derivative such as L-ascorbic acid or sodium L-ascorbate, an
ascorbic acid preparation obtained by coating ascorbic acid with an
emulsifier or the like, and a mixture containing two or more of
those vitamin Cs at an arbitrary rate. In addition, natural
products containing vitamin C such as acerola or lemon may also be
used.
Examples of the iron preparation include: an inorganic iron such as
ferrous sulfate, sodium ferrous citrate, or ferric pyrophosphate;
an organic iron such as heme iron, ferritin iron, or lactoferrin
iron; and a mixture containing two or more of those irons at an
arbitrary rate. In addition, natural products containing iron such
as spinach or liver may also be used. Moreover, examples of the
collagen include: an extract obtained by treating bone, skin, or
the like of a mammal such as bovine or swine with an acid or
alkaline; a peptide obtained by hydrolyzing the extract with a
protease such as pepsin, trypsin, or chymotrypsin; and a mixture
containing two or more of those collagens at an arbitrary rate.
Collagens extracted from plant sources may also be used.
(ii) Skin Disorders
The silicone-based biophotonic compositions and methods of the
present disclosure may be used in a treatment of a skin disorder
that may include, but is not limited to, erythema, telangiectasia,
actinic telangiectasia, basal cell carcinoma, contact dermatitis,
dermatofibrosarcoma protuberans, genital warts, hidradenitis
suppurativa, melanoma, merkel cell carcinoma, nummular dermatitis,
molloscum contagiosum, psoriasis, psoriatic arthritis, rosacea,
scabies, scalp psoriasis, sebaceous carcinoma, squamous cell
carcinoma, seborrheic dermatitis, seborrheic keratosis, shingles,
tinea versicolor, warts, skin cancer, pemphigus, sunburn,
dermatitis, eczema, rashes, impetigo, lichen simplex chronicus,
rhinophyma, perioral dermatitis, pseudofolliculitis barbae,
erythema multiforme, erythema nodosum, granuloma annulare, actinic
keratosis, purpura, alopecia areata, aphthous stomatitis, drug
eruptions, dry skin, chapping, xerosis, ichthyosis vulgaris, fungal
infections, herpes simplex, intertrigo, keloids, keratoses, milia,
moluscum contagiosum, pityriasis rosea, pruritus, urticaria, and
vascular tumors and malformations. Dermatitis includes contact
dermatitis, atopic dermatitis, seborrheic dermatitis, nummular
dermatitis, generalized exfoliative dermatitis, and statis
dermatitis. Skin cancers include melanoma, basal cell carcinoma,
and squamous cell carcinoma.
(iii) Acne and Acne Scars
The silicone-based biophotonic compositions and methods of the
present disclosure may be used to treat acne. As used herein,
"acne" means a disorder of the skin caused by inflammation of skin
glands or hair follicles. The silicone-based biophotonic
compositions and methods of the disclosure can be used to treat
acne at early pre-emergent stages or later stages where lesions
from acne are visible. Mild, moderate and severe acne can be
treated with embodiments of the silicone-based biophotonic
compositions and methods. Early pre-emergent stages of acne usually
begin with an excessive secretion of sebum or dermal oil from the
sebaceous glands located in the pilosebaceous apparatus. Sebum
reaches the skin surface through the duct of the hair follicle. The
presence of excessive amounts of sebum in the duct and on the skin
tends to obstruct or stagnate the normal flow of sebum from the
follicular duct, thus producing a thickening and solidification of
the sebum to create a solid plug known as a comedone. In the normal
sequence of developing acne, hyperkeratinazation of the follicular
opening is stimulated, thus completing blocking of the duct. The
usual results are papules, pustules, or cysts, often contaminated
with bacteria, which cause secondary infections. Acne is
characterized particularly by the presence of comedones,
inflammatory papules, or cysts. The appearance of acne may range
from slight skin irritation to pitting and even the development of
disfiguring scars. Accordingly, the silicone-based biophotonic
compositions and methods of the present disclosure can be used to
treat one or more of skin irritation, pitting, development of
scars, comedones, inflammatory papules, cysts, hyperkeratinazation,
and thickening and hardening of sebum associated with acne.
Some skin disorders present various symptoms including redness,
flushing, burning, scaling, pimples, papules, pustules, comedones,
macules, nodules, vesicles, blisters, telangiectasia, spider veins,
sores, surface irritations or pain, itching, inflammation, red,
purple, or blue patches or discolorations, moles, and/or
tumors.
The silicone-based biophotonic compositions and methods of the
present disclosure may be used to treat various types of acne. Some
types of acne include, for example, acne vulgaris, cystic acne,
acne atrophica, bromide acne, chlorine acne, acne conglobata, acne
cosmetica, acne detergicans, epidemic acne, acne estivalis, acne
fulminans, halogen acne, acne indurata, iodide acne, acne keloid,
acne mechanica, acne papulosa, pomade acne, premenstral acne, acne
pustulosa, acne scorbutica, acne scrofulosorum, acne urticata, acne
varioliformis, acne venenata, propionic acne, acne excoriee, gram
negative acne, steroid acne, and nodulocystic acne.
In certain embodiments, the silicone-based biophotonic compositions
of the present disclosure is used in conjunction with systemic or
topical antibiotic treatment. For example, antibiotics used to
treat acne include tetracycline, erythromycin, minocycline,
doxycycline, which may also be used with the compositions and
methods of the present disclosure. The use of the silicone-based
biophotonic composition can reduce the time needed for the
antibiotic treatment or reduce the dosage.
(iv) Wound Healing
The silicone-based biophotonic compositions and methods of the
present disclosure may be used to treat wounds, promote wound
healing, and promote tissue. Wounds that may be treated by the
silicone-based biophotonic compositions and methods of the present
disclosure include, for example, injuries to the skin and
subcutaneous tissue initiated in different ways (e.g., pressure
ulcers from extended bed rest, wounds induced by trauma or surgery,
burns, ulcers linked to diabetes or venous insufficiency, wounds
induced by conditions such as periodontitis) and with varying
characteristics. In certain embodiments, the present disclosure
provides silicone-based biophotonic compositions and methods for
treating and/or promoting the healing of, for example, burns,
incisions, excisions, lesions, lacerations, abrasions, puncture or
penetrating wounds, surgical wounds, contusions, hematomas,
crushing injuries, amputations, sores and ulcers.
Silicone-based biophotonic compositions and methods of the present
disclosure may be used to treat and/or promote the healing of
chronic cutaneous ulcers or wounds, which are wounds that have
failed to proceed through an orderly and timely series of events to
produce a durable structural, functional, and cosmetic closure. The
vast majority of chronic wounds can be classified into three
categories based on their etiology: pressure ulcers, neuropathic
(diabetic foot) ulcers and vascular (venous or arterial)
ulcers.
For example, the present disclosure provides silicone-based
biophotonic compositions and methods for treating and/or promoting
healing of a diabetic ulcer. Diabetic patients are prone to foot
and other ulcerations due to both neurologic and vascular
complications. Peripheral neuropathy can cause altered or complete
loss of sensation in the foot and/or leg. Diabetic patients with
advanced neuropathy lose all ability for sharp-dull discrimination.
Any cuts or trauma to the foot may go completely unnoticed for days
or weeks in a patient with neuropathy. A patient with advanced
neuropathy loses the ability to sense a sustained pressure insult,
as a result, tissue ischemia and necrosis may occur leading to for
example, plantar ulcerations. Microvascular disease is one of the
significant complications for diabetics which may also lead to
ulcerations. In certain embodiments, silicone-based biophotonic
compositions and methods of treating a chronic wound are provided
here in, where the chronic wound is characterized by diabetic foot
ulcers and/or ulcerations due to neurologic and/or vascular
complications of diabetes.
In other examples, the present disclosure provides silicone-based
biophotonic compositions and methods for treating and/or promoting
healing of a pressure ulcer. Pressure ulcers include bed sores,
decubitus ulcers and ischial tuberosity ulcers and can cause
considerable pain and discomfort to a patient. A pressure ulcer can
occur as a result of a prolonged pressure applied to the skin.
Thus, pressure can be exerted on the skin of a patient due to the
weight or mass of an individual. A pressure ulcer can develop when
blood supply to an area of the skin is obstructed or cut off for
more than two or three hours. The affected skin area can turn red,
become painful and necrotic. If untreated, the skin can break open
and become infected. A pressure ulcer is therefore a skin ulcer
that occurs in an area of the skin that is under pressure from e.g.
lying in bed, sitting in a wheelchair, and/or wearing a cast for a
prolonged period of time. Pressure ulcers can occur when a person
is bedridden, unconscious, unable to sense pain, or immobile.
Pressure ulcers often occur in boney prominences of the body such
as the buttocks area (on the sacrum or iliac crest), or on the
heels of foot.
There are three distinct phases in the wound healing process.
First, in the inflammatory phase, which typically occurs from the
moment a wound occurs until the first two to five days, platelets
aggregate to deposit granules, promoting the deposit of fibrin and
stimulating the release of growth factors. Leukocytes migrate to
the wound site and begin to digest and transport debris away from
the wound. During this inflammatory phase, monocytes are also
converted to macrophages, which release growth factors for
stimulating angiogenesis and the production of fibroblasts.
Second, in the proliferative phase, which typically occurs from two
days to three weeks, granulation tissue forms, and
epithelialization and contraction begin. Fibroblasts, which are key
cell types in this phase, proliferate and synthesize collagen to
fill the wound and provide a strong matrix on which epithelial
cells grow. As fibroblasts produce collagen, vascularization
extends from nearby vessels, resulting in granulation tissue.
Granulation tissue typically grows from the base of the wound.
Epithelialization involves the migration of epithelial cells from
the wound surfaces to seal the wound. Epithelial cells are driven
by the need to contact cells of like type and are guided by a
network of fibrin strands that function as a grid over which these
cells migrate. Contractile cells called myofibroblasts appear in
wounds, and aid in wound closure. These cells exhibit collagen
synthesis and contractility, and are common in granulating
wounds.
Third, in the remodeling phase, the final phase of wound healing
which can take place from three weeks up to several years, collagen
in the scar undergoes repeated degradation and re-synthesis. During
this phase, the tensile strength of the newly formed skin
increases.
However, as the rate of wound healing increases, there is often an
associated increase in scar formation. Scarring is a consequence of
the healing process in most adult animal and human tissues. Scar
tissue is not identical to the tissue which it replaces, as it is
usually of inferior functional quality. The types of scars include,
but are not limited to, atrophic, hypertrophic and keloidal scars,
as well as scar contractures. Atrophic scars are flat and depressed
below the surrounding skin as a valley or hole. Hypertrophic scars
are elevated scars that remain within the boundaries of the
original lesion, and often contain excessive collagen arranged in
an abnormal pattern. Keloidal scars are elevated scars that spread
beyond the margins of the original wound and invade the surrounding
normal skin in a way that is site specific, and often contain
whorls of collagen arranged in an abnormal fashion.
In contrast, normal skin consists of collagen fibers arranged in a
basket-weave pattern, which contributes to both the strength and
elasticity of the dermis. Thus, to achieve a smoother wound healing
process, an approach is needed that not only stimulates collagen
production, but also does so in a way that reduces scar
formation.
Certain embodiments of the silicone-based biophotonic compositions
and methods of the present disclosure may promote wound healing by
promoting the formation of substantially uniform epithelialization;
promoting collagen synthesis; promoting controlled contraction;
and/or by reducing the formation of scar tissue. In certain
embodiments, the biophotonic compositions and methods of the
present disclosure may promote wound healing by promoting the
formation of substantially uniform epithelialization. In some
embodiments, the silicone-based biophotonic compositions and
methods of the present disclosure may modulate or promote collagen
synthesis. In some other embodiments, the silicone-based
biophotonic compositions and methods of the present disclosure may
promote controlled contraction. In certain embodiments, the
silicone-based biophotonic compositions and methods of the present
disclosure may promote wound healing, for example, by reducing the
formation of scar tissue.
In the methods of the present disclosure, the silicone-based
biophotonic compositions of the present disclosure may also be used
in combination with negative pressure assisted wound closure
devices and systems.
In certain embodiments, the silicone-based biophotonic composition
is kept in place for up to one, two or 3 weeks, and illuminated
with light which may include ambient light at various intervals. In
this case, the silicone-based biophotonic composition may be
covered up in between exposure to light with an opaque material or
left exposed to light.
(6) Kits
The present disclosure also provides kits for preparing a
silicone-based biophotonic compositions and/or providing any of the
components required for forming silicone-based biophotonic
compositions of the present disclosure.
In some embodiments, the kit includes containers comprising the
components or compositions that can be used to make the
silicone-based biophotonic compostions of the present disclosure.
In some embodiments, the kit includes the silicone-based
biophotonic composition of the present disclosure. The different
components making up the silicone-based biophotonic compositions of
the present disclosure may be provided in separate containers. For
example, the surfactant phase may be provided in a container
separate from the silicone phase. Examples of such containers are
dual chamber syringes, dual chamber containers with removable
partitions, sachets with pouches, and multiple-compartment blister
packs. Another example is one of the components being provided in a
syringe which can be injected into a container of another
component.
In other embodiments, the kit comprises a systemic drug for
augmenting the treatment of the silicone-based biophotonic
composition of the present disclosure. For example, the kit may
include a systemic or topical antibiotic, hormone treatment (e.g.
for acne treatment or wound healing), or a negative pressure
device.
In other embodiments, the kit comprises a means for mixing or
applying the components of the silicone-based biophotonic
compositions.
In certain embodiments of the kit, the kit may further comprise a
light source such as a portable light with a wavelength appropriate
to activate the chromophore of the silicone-based biophotonic
composition. The portable light may be battery operated or
re-chargeable.
Written instructions on how to use the silicone-based biophotonic
compositions in accordance with the present disclosure may be
included in the kit, or may be included on or associated with the
containers comprising the silicone-based biophotonic composition or
the components making up the silicone-based biophotonic
compositions of the present disclosure.
Identification of equivalent silicone-based biophotonic
compositions, methods and kits are well within the skill of the
ordinary practitioner and would require no more than routine
experimentation, in light of the teachings of the present
disclosure.
Variations and modifications will occur to those of skill in the
art after reviewing this disclosure. The disclosed features may be
implemented, in any combination and sub-combinations (including
multiple dependent combinations and sub-combinations), with one or
more other features described herein. The various features
described or illustrated above, including any components thereof,
may be combined or integrated in other systems. Moreover, certain
features may be omitted or not implemented. Examples of changes,
substitutions, and alterations are ascertainable by one skilled in
the art and could be made without departing from the scope of the
information disclosed herein. All references cited herein are
incorporated by reference in their entirety and made part of this
application.
Practice of the disclosure will be still more fully understood from
the following examples, which are presented herein for illustration
only and should not be construed as limiting the disclosure in any
way.
EXAMPLES
Example 1: Silicone-Based Biophotonic Composition (25%
Pluronic-F127)
Preparation of 25% Wt % Pluronic-F127 Solutions (Surfactant
Phase)
Typical preparation of thermogelling solutions of Pluronic
comprised dissolving a measured mass of Pluronic F-127 in a
measured volume of cold, de-ionised water (.about.4.degree. C.).
The concentration of Pluronic is expressed in weight per volume of
H.sub.2O.
Thus, for the preparation of a stock thermogelling Pluronic
solution (25% w/v), a mass of 25.00 g of Pluronic F-127 was added,
under magnetic stirring, to 100 mL of H.sub.2O in an Erlenmeyer
flask of 250 mL. The Erlenmeyer with the solution was then cooled
in an ice bath (between 2 and 4.degree. C.), while continuing
stirring for about 1 hour, until complete dissolution of the
Pluronic F-127. The resulting solution was then stored in the
fridge at about 4.degree. C.
A gelation test was performed which indicated that the solution
formed into a hydrogel after approximately 5 minutes at room
temperature (.about.22.degree. C.).
Preparation of Silicone-15/85 (Silicone Phase)
A silicone-15/85 component for the silicone-based biophotonic was
prepared by mixing 15% of Sylgard-184 elastomer kit and 85% of
Sylgard-527 gel kit. Thus, typical mixture of silicone-15/85 was
prepared by thoroughly mixing 2.667 g of Sylgard-184 (composed of
2.423 g of part A of the Sylgard-184 kit and 0.244 g of part B of
the Sylgard-184 kit), with 15.151 g of Sylgard-527 (composed of
7.574 g of part A of the Sylgard-527 kit and 7.577 g of part B of
the Sylgard-527 kit). The silicone-15/85 mixture was cooled down to
-4.degree. C. in order to maintain it in a liquid form.
Preparation of the Silicone-Based Biophotonic Composition
To form the silicone-based biophotonic composition, 2.0 mL of the
cold Pluronic-F127 themogelling solution containing 0.327 mg of
Eosin Y and 0.327 g of Fluorescein was added to 9.221 g of the
silicone-15/85 mixture, freshly prepared, under vigorous stirring
in order to create an extremely fine emulsion. Thereafter, in order
to form a silicone-based biophotonic membrane, the resulting
mixture was cast onto petri dishes. The cast amount was controlled
so as to obtain a membrane thickness of 2 mm. The casted,
silicone-based biophotonic membrane mixture was then cured for 5
hours at 40.degree. C. and under humid atmosphere in an
incubator.
The emulsion that was formed on completion of the mixing of the
surfactant phase and the silicone phase was a very fine and
highly-stable micro-emulsion or gel. Without being bound to a
particular theory, it was thought that these properties of the
micro-emulsion may have resulted from the hydrophobic nature of
silicone and the surfactant properties of Pluronic-F127 When cast
in the Pertri dish and after curing, the resulting silicone-based
biophotonic membrane was homogeneous and flexible. The membrane was
thereafter tested to evaluate whether the chromophores (Eosin Y and
fluorescein) might leach from the silicone-based biophotonic
membrane, as sample of the membrane was immersed in a
phosphate-buffered saline (PBS) solution for 24 hours and no
leaching of the chromophores was observed
In a second experiment, 0.75 mL of Pluronic-F127 thermogelling
solution containing 0.123 mg of Eosin Y and 0.123 mg of fluorescein
was added to 6.744 g of silicone-15/85 (prepared as described
above) under vigorous stirring. The resultant uniform microemulsion
was extremely fine and showed high stability. Aliquots of the
micro-emulsion were casted onto petri dishes so as to obtain a
thickness of 2 mm, then cured for 5 hours at 40.degree. C. and
under humid atmosphere in an incubator.
Light emitted through and by a silicone-based biophotonic membrane
prepared from this second experiment was measured using a SP-100
spectroradiometer (SP-100, ORB Optronix) whilst being illuminated
with light having a peak emission wavelength of 450 nm (peak
wavelength ranging between 400-470 nm and a power density of about
30-150 mW/cm.sup.2) for 5 minutes. As can be seen in FIGS. 1-4, the
chromophores did not fully photobleach after 15 minutes of
illumination in 5 minute intervals.
Example 2--Cytokines and Growth Factors in DHF
In order to gain a more detailed picture of the biological effect
mediated by the silicone-based biophotonic membrane of Example 1
(second experiment), Human Cytokine Antibody Array (RayBio
C-Series, RayBiotech, Inc.) was performed. Cytokines broadly
defined as secreted cell-cell signaling proteins play important
roles in inflammation, innate immunity, apoptosis, angiogenesis,
cell growth and differentiation. Simultaneous detection of multiple
cytokines provides a powerful tool to study cell activity.
Regulation of cellular processes by cytokines is a complex, dynamic
process, often involving multiple proteins. Positive and negative
feedback loops, pleiotrophic effects and redundant functions,
spatial and temporal expression of or synergistic interactions
between multiple cytokines, even regulation via release of soluble
forms of membrane-bound receptors, are all common mechanisms
modulating the effects of cytokine signaling.
DHF (Derman Human Fibroblast) and THP1(human acute monocytic
leukemia cells) were used as an in vitro model to study the effect
of the blue light in combination with the light emitted by the
silicone-based biophotonic membrane on the secretion of the
inflammatory cytokines, chemokines and growth factors. Excessive,
uncontrolled inflammation is detrimental to the host and can impair
wound healing processes amongst other things. The purpose of this
study was to demonstrate that blue light in combination with the
fluorescence emitted by the silicone-based biophotonic membrane(s)
is able to down-regulate the production of pro-inflammatory
cytokines and chemokines and improve/accelerate the healing
process.
Briefly, a non-toxic concentration of TGF (3-1 was used to
stimulate DHF cells, and IFN.gamma. and LPS were used to stimulate
PMA-treated THP-1 cells. The membrane of Example 1 (second
experiment) was then positioned 5 cm above the cell cultures and
illuminated with blue light (450 nm).
Cell culture mediums were collected 24 h post-illumination and
incubated with arrayed antibody membranes according to manufacturer
instructions (Human Cytokine Antibody Array, RayBio C-series from
Raybiotech). Signals were quantified with Image J.RTM. software.
For each experiment, the XTT assay (cell viability assay) was
performed to normalize the quantity of cytokine secreted to the
cell viability (in all cases the viability was over 90% showing a
non-toxic effect of the treatment). All samples were done in
quadruplets.
The effect of illuminated membrane on cytokines and growth factor
secretion in DHF and THP-1 cells is summarized in the Tables 1 and
2 below.
TABLE-US-00001 TABLE 1 Modulation of protein expression in Dermal
Human Fibroblasts activated by TGFB1 24 hours after treatment with
blue light + silicone- based biophotonic membrane compared to
control untreated cells. Silicone- basedbipohotonic membrane
Cytokines IL2 -- IL3 .dwnarw..dwnarw..dwnarw. IL4 IL6 -- IL8
.dwnarw..dwnarw..dwnarw. IL10 IL12 p40/70 -- IL13 .dwnarw. IL15
TNF-alpha .dwnarw. TNF-beta .dwnarw..dwnarw..dwnarw. IL1-alpha
.dwnarw..dwnarw..dwnarw. IL1-beta .dwnarw..dwnarw. IFN-gamma
.dwnarw..dwnarw..dwnarw. MCP1 .dwnarw..dwnarw..dwnarw. MCP2
.dwnarw..dwnarw..dwnarw. MCP3 M-CSF .dwnarw..dwnarw..dwnarw. MDC --
MIG MIP-1-delta .dwnarw..dwnarw..dwnarw. RANTES
.dwnarw..dwnarw..dwnarw. TARC Growth Factors EGF -- IGF-1 ANG VEGF
PDGF-BB .dwnarw..dwnarw..dwnarw. ENA-78 .dwnarw..dwnarw..dwnarw.
G-CSF GM-CSF .dwnarw..dwnarw..dwnarw. GRO .dwnarw..dwnarw..dwnarw.
GRO-alpha .dwnarw..dwnarw..dwnarw. TGFbeta1
.dwnarw..dwnarw..dwnarw. Leptin -- .dwnarw. less than 25% decrease
.dwnarw..dwnarw. 25-50% decrease .dwnarw..dwnarw..dwnarw. more than
50% decrease -- No modulation less than 25% increase 25-50%
increase more than 50% increase
TABLE-US-00002 TABLE 2 Modulation of protein expression in THP1
cells differentiated into macrophages 24 hours after treatment with
blue light + silicone- based biophotonic membrane compared to
control untreated cells. Silicone- based biophotonic membrane
Cytokines IL2 -- IL3 .dwnarw..dwnarw..dwnarw. IL4 -- IL6
.dwnarw..dwnarw..dwnarw. IL8 .dwnarw..dwnarw. IL10 IL12 p40/70 --
IL13 -- IL15 TNF-alpha .dwnarw..dwnarw..dwnarw. TNF-beta --
IL1-alpha .dwnarw. IL1-beta .dwnarw..dwnarw..dwnarw. IFN-gamma --
MCP1 MCP2 .dwnarw..dwnarw..dwnarw. MCP3 -- M-CSF -- MDC -- MIG --
MIP-1-delta -- RANTES .dwnarw..dwnarw. TARC Growth Factors EGF --
IGF-1 -- ANG -- VEGF PDGF-BB ENA-78 -- G-CSF -- GM-CSF
.dwnarw..dwnarw..dwnarw. GRO .dwnarw..dwnarw. GRO-alpha .dwnarw.
TGFbeta1 -- Leptin .dwnarw. less than 25% decrease .dwnarw..dwnarw.
25-50% decrease .dwnarw..dwnarw..dwnarw. more than 50% decrease --
No modulation less than 25% increase 25-50% increase more than 50%
increase
Results from the cytokine/chemokine array assay revealed that the
treatment with the silicone-based biophotonic membrane of Example 1
negatively modulated pro-inflammatory cytokines (such as TNF alpha,
IL-6, IL-8, IL-1 alpha, IL-1 beta, IFN.gamma.) and pro-inflammatory
chemokines (such as MCP-1, -2, RANTES, GRO,) production. The
results also indicated that the treatment utilizing the
silicone-based biophotonic membrane demonstrated an ability to
negatively modulate growth factors secretion (such as TGF-beta1,
and PDGF-BB) in DHF cells.
Example 3--Proliferation Level in DHF Cells Upon Illumination by a
Silicone-Based Biophotonic Membrane
In order to gain more detailed picture of the biological effect
mediated by the silicone-based biophotonic membrane of Example 1
(second experiment) and its implication in a wound healing process,
cellular proliferation was assessed in Human Dermal Fibroblast
(DHF) experimental system. In tissues, within four-five days upon
injury, matrix-generating cells i.e. fibroblasts, move into the
granulation tissue. Their migration to and proliferation within the
wound site are prerequisites for wound granulation and consecutive
healing. Fibroblasts then participate in the construction of scar
tissue and its remodeling. Thus viable, actively dividing
fibroblast are crucial player in healing progression.
The present experiment utilized an XTT assay to measure cell
viability. The XTT-based method measures the mitochondrial
dehydrogenase activity of proliferating cells. In brief, the
mitochondrial dehydrogenases of viable cells reduce the tetrazolium
ring of XTT, yielding an orange derivative, which is water soluble.
The absorbance of the resulting orange solution is measured
spectrophotometrically. An increase or decrease in cells number
relative to control cells, results in an accompanying change in the
amount of orange derivative, indicating the changes in the number
of viable, dividing cells.
DHF cells were illuminated for 5 min with the silicone-based
biophotonic membrane of Example 1. 24 h post-treatment XTT solution
was added to the cells. Four hours later the absorbance of orange
supernatant was measured spectrophotometrically. The difference in
the number of actively proliferating fibroblasts as compared to
non-illuminated control was calculated.
The XTT assay showed that the silicone membrane of Example 1 did
not modulate DHF proliferation under the test conditions as
compared to a control (non-treated cells).
Example 4--Evaluation of a Silicone-Based Biophotonic Thermogel of
the Present Description for a Prevention of Scarring
Hypertrohic scars (HTS) result from excessive dermal fibrosis
involving myofibroblasts. They occur after an injury to the dermis.
In addition to their disfiguring characteristic, scars can be
itchy, rigid and painful. Excessive production of collagen and
other extracellular matrix (ECM) proteins and/or deficient
degradation and remodeling of ECM are the main causes of scar
formation. These phenomenon occur when the inflammatory response to
injury is prolonged. In HTS, the growth factors, TGF.beta.1 and
PDGF are over expressed by fibroblasts. They are major proteins in
HTS (Avouac J, et al. Inhibition of activator protein 1 signaling
abrogates transforming growth factor b-mediated activation of
fibroblasts and prevents experimental fibrosis. Arthritis
Rheumatism, 2012, volume 64:1642-4652; Trojanowska M, Role of PDGF
in fibrotic diseases and systemic sclerosis. Rheumatology, 2008,
volume 47: v2-v4). TGFb1 is responsible for the excessive collagen
secretion and the reduction of matrix metalloproteinases (MMPs)
such as collagenase (Cutroneo K R. TGF-beta-induced fibrosis and
SMAD signaling: oligo decoys as natural therapeutics for inhibition
of tissue fibrosis and scarring., Wound Rep Regen 2007, volume 15:
S54-60; Chen Z C, Raghunath M. Focus on collagen: In vitro systems
to study fibrogenesis and antifibrosis--state of the art.
Fibrogenesis Tissue Repair, 2009, volume 2: 7). PDGF is a potent
chemoattractant for fibroblasts and constitutes a good target for
the treatment of fibrosis (Beyer C, Distler J H W. Tyrosine kinase
signaling in fibrotic disorders. Translation of basic research to
human disease. Biochem Biophys Acta, 2013, volume 1832: 897-904).
HTS have high expression of MMP-2 and low expression of MMP-9
(Gauglitz G G et al. Hypertrophic scarring and keloids:
pathomechanisms and current and emerging treatment strategies. Mol
Med, 2011; volume 17: 113-125).
Experimental Design
a) Protein Secretion-Inflammatory Mediators, Cytokines, Growth
Factors
A Dermal Human Fibroblasts (DHF) cell culture model was used as in
vitro model to study the effect of a treatment comprising of an
illumination with an actinic light source emitting a non-coherent
blue light upon a silicone-based biophotonic membrane containing
the chromophores Eosin Y and fluorescein may have on the secretion
of various proteins that function as inflammatory mediators, or
growth factors, or which are involved in tissue remodeling (such as
matrix metalloproteinases (MMPs), and tissue inhibitors of matrix
metalloproteinases (TIMPs).
For this experimental model, the cells were illuminated for a
period of 5 minutes using the above-described silicone-based
biophotonic membrane together with a visible blue light (KLOX
Multi-LED light) at the distance of 5 cm. The blue light and
fluorescence dose received by the cells during the illumination
time are presented in Table 3.
TABLE-US-00003 TABLE 3 Dose (J/cm2) of blue light and fluorescence
received by the cells during 5 minutes illumination Purple 10.95
Blue 6.33 Green 0.53 Yellow 0.25 Orange 0.15 Red 0.16 Total J/cm2
18.37
DHF cells were cultured on a glass bottom dish (approximately 2 mm
thickness). One hour prior to illumination, the cells were treated
with non-toxic concentration of TGF.beta.1 (5 ng/ml) to induce the
hyperproliferative state that is that is typically observed in the
process of the formation of hypertrophic scars. TGF.beta.1 was
maintained in the culture medium after the illumination to mimic
the scarring condition through whole time during which the assay
was performed. The silicone-based biophotonic membrane as described
above was applied on the other side of the glass dish (i.e. on the
exterior surface of the dish) and illuminated at 5 cm distance
using blue visible light (KLOX Thera.TM. lamp). Cells were also
treated with light alone, which served as an internal control to
ensure that the combination of light with the silicone-based
biophotonic membrane containing the Eosin Y and fluorescein
chromophores exerted a biological effect compared to light alone.
At 24-hours post-treatment, the supernatant was collected and
arrays were performed to evaluate the inflammatory cytokines,
chemokines and growth factors production profile resulting from the
treatment. The lists of proteins analyzed for each antibody array
are presented below in Tables 4 and 5.
Antibodies Array profiles
TABLE-US-00004 TABLE 4 Human Cytokine Antibody Array C3 A B C D E F
G H I J K L 1 POS POS NEG NEG ENA-78 G-CSF GM-CSF GRO GRO I-309
IL-1 IL-1 alpha alpha beta 2 POS POS NEG NEG ENA-78 G-CSF GM-CSF
GRO GRO I-309 IL-1 IL-1 alpha alpha beta 3 IL-2 IL-3 IL-4 IL-5 IL-6
IL-7 IL-8 IL-10 IL-12 IL-13 IL-15 IFN p40/70 gamma 4 IL-2 IL-3 IL-4
IL-5 IL-6 IL-7 IL-8 IL-10 IL-12 IL-13 IL-15 IFN p40/70 gamma 5
MCP-1 MCP-2 MCP-3 M-CSF MDC MIG MIP-1 RANTES SCF SDF-1 TARC TGF
delta beta 1 6 MCP-1 MCP-2 MCP-3 M-CSF MDC MIG MIP-1 RANTES SCF
SDF-1 TARC TGF delta beta 1 7 TNF TNF EGF IGF-1 ANG OSM THPO VEGF
PDGF Leptin NEG POS alpha beta BB 8 TNF TNF EGF IGF-1 ANG OSM THPO
VEGF PDGF Leptin NEG POS alpha beta BB POS = Positive Control Spot
NEG = Negative Control Spot BLANK = Blank Spot
TABLE-US-00005 TABLE 5 Human Growth Factor Antibody Array C1 A B C
D E F G H I J K L 1 POS POS NEG NEG AREG bFGF b-NGF EGF EGFR FGF-4
FGF-6 FGF-7 2 POS POS NEG NEG AREG bFGF b-NGF EGF EGFR FGF-4 FGF-6
FGF-7 3 G-CSF GDNF GM HB HGF IGFBP1 IGFBP2 IGFBP3 IGFBP4 IGFBP6
IGF-1 IGF-1 CSF EGF sR 4 G-CSF GDNF GM HB HGF IGFBP1 IGFBP2 IGFBP3
IGFBP4 IGFBP6 IGF-1 IGF-1 CSF EGF sR 5 IGF-2 M-CSF M-CSF R NT-3
NT-4 PDGF R PDGF R PDGF PDGF PDGF PLGF SCF alpha beta AA AB BB 6
IGF-2 M-CSF M-CSF R NT-3 NT-4 PDGF R PDGF R PDGF PDGF PDGF PLGF SCF
alpha beta AA AB BB 7 SCF R TGF TGF TGF TGF VEGF VEGF VEGF VEGF D
BLANK BLANK POS alpha beta beta 2 beta 3 R2 R3 8 SCF R TGF TGF TGF
TGF VEGF VEGF VEGF VEGF D BLANK BLANK POS alpha beta beta 2 beta 3
R2 R3 POS = Positive Control Spot NEG = Negative Control Spot BLANK
= Blank Spot
To assess the potential cytotoxicity of the treatment, supernatants
from the treated cell cultures were also screened for lactate
dehydrogenase (LDH) activity. LDH is an intracellular enzyme that
is released in the culture medium when the cell is damaged. It is a
marker of cytotoxicity. The assay quantifies the LDH activity that
reduces NAD to NADH. NADH is specifically detected by
colorimetry.
b) Cell Proliferation (DHF Cell Cultures)
Prior to the treatment, cells were subjected to starvation
conditions (medium deprived of serum and hormones) in order to be
synchronised in G1 phase. Following synchronisation, the DHF were
subjected to the treatment comprising the silicone-based
biophotonic thermogel and blue light illumination (with the
intensity of 14.4 J/cm.sup.2 at 5 cm distance). Cells were
monitored for their proliferation at 24 h, 48 h, and 72 h
post-treatment using CyQUANT direct cell proliferation assay.
c) In vivo Study Using a Dermal Fibrotic Mouse--Human Skin Graft
Model System
To evaluate the potential of the silicone-based biophotonic
composition treatment of the present disclosure to promote wound
healing and prevent scarring, an in vivo mouse model system was
utilized, more particularly, a dermal fibrotic mouse model, in
which the split thickness human skin transplanted to full thickness
excision wounds on the back of nude mouse developed a thickened,
raised, contracted scar resembling human HTS (see Montazi M et al.
A nude mouse model of hypertrophic scar shows morphologic and
histologic characteristics of human hypertrophic scar. Wound Rep
Reg, 2013, volume 21: 77-87).
To evaluate the treatment comprising the silicone-based biophotonic
composition (containing Eosin Y and fluorescein) (prepared as per
Example 1, experiment number 2 described above) and the visible
blue light (KLOX Multi LED light) illumination, the biophotonic
composition-light illumination treatment was applied (either in the
form of an unpolymerized gel or as a polymerized membrane) using
the illumination times and distance as described for the in vitro
experiments of this Example 4, however in this in vivo system the
biophonic composition was applied to be in topical (physical)
contact) to the skin grafted wounds. Treatment with the
light-biophotonic composition began at day 7 post-transplantation
with the mice being under a light general anesthesia via halothane
nasal application. The treatment was done twice per week for a
period of 3 weeks. The animals were sacrificed one week after the
last treatment. Control animals did not receive the treatment and
another group received the blue light only. The wounds were
monitored by digital photography weekly before the animals were
euthanized at the 4 weeks post-treatment point and the excised
xenografts were examined.
The quantification of scar thickness and vascularity were done on
hematoxylin & eosin (H&E) stained section images. Using
Image J, the measurements of dermal thickness were done in high
power images, with the dermal thickness being the distance between
the epidermal-dermal junction and the dermal-adipose layer
junction. Three measurements were taken per sample. The degree of
vascularity was assessed by counting the number of blood vessels in
five high power fields (HPFs) of the dermis.
Masson's Trichrome staining (as known in the art) was used to
detect collagen fibers in the dermis. Using polarized light
microscopy to examine the stained specimens, collagen fibers could
be observed as being green in color, while nuclei appeared in black
and cytoplasm and keratin in red.
Results
a) Effect of a Silicone-Based Biophotonic Membrane with Blue Light
Illumination Treatment on Production of Inflammatory Mediators
Production in DHF Cells
At 24 h post-treatment supernatant was collected and inflammatory
cytokine array was performed to evaluate the inflammatory cytokines
production profile upon silicone-based biophotonic membrane
(containing Eosin Y and fluorescein) treatment in combination with
KLOX Multi-LED light. The results of the array are summarized in
Table 6.
Analysis of LDH activity showed that no significant cytotoxic
effect of the treatment was observed in all of the silicone-based
biophotonic membrane illuminated samples.
TABLE-US-00006 TABLE 6 Summary of significant up (.uparw.) and
down-regulation (.dwnarw.) observed in inflammatory mediators
production (cytokines in red, chemokines in blue) and growth
factors (in black) compared to non-treated controls. Decrease
Increase IL-3, IL-8, TNF.beta., IL-1.alpha., IL-1.beta., IL-10,
MIG, IGF1, ANG, VEGF, G-CSF IFN.gamma., MCP1 MCP2, M-CSF,
MIP1.delta., RANTES, GRO, GRO.alpha. PDGF-BB, ENA-78, GM-CSF,
TGF.beta.1
PDGF-BB and TGFb1 and important growth factors implicated in the
pathogenesis of scarring. The ability of the treatment to decrease
significantly these factors is beneficial. Furthermore, a number of
pro-inflammatory mediators were also observed to be decreased in
the cells subjected to the treated versus control cells, while it
was observed that certain anti-inflammatory cytokines were
increased in the treated cells, for example IL-10.
b) Cell Proliferation (DHF Cell Cultures)
In reference to the data regarding growth factors induced upon the
silicone-based biophotonic membrane of the present disclosure, as
can be seen in Table 6 (above), the induced growth factors are
mostly involved in blood vessel formation as opposed to being
growth factors that are involved in cell proliferation.
Furthermore, results from the cell proliferation assay performed in
this Example 4 also show that the silicone-based biophotonic
membrane did not induce cell proliferation. This lack of effect on
fibroblast proliferation can be considered to be beneficial in
hypertrophic scarring, given that hypertrophic scarring is
characterized as a hyperproliferative disorder.
c) In vivo Study Using a Dermal Fibrotic Mouse--Human Skin Graft
Model System
Morphologically, there were no visible significant differences
between the groups grossly or in wound contraction measured by
planimetery. However, by 4 weeks post-engraftment, significant
reductions in scar thickness were measured histologically in the
silicone-based biophotonic composition (applied as an unpolymerized
gel)-plus-light and in the silicone-based biophotonic
membrane-plus-light treatment groups as compared to both the
control and light-only groups, (1.35.+-.0.07, 1.35.+-.0.08 vs
1.69.+-.0.13, 2.07.+-.0.08 mm P<0.05) with improvements in
re-epithelialization. These results are also presented in a
graphical format in FIG. 5.
With respect to the effect of the treatments using the
silicone-based biophotonic composition, morphological improvements
in collagen fiber bundles and orientation (based on Masson
Trichrome staining) were associated with accelerated collagen
remodeling in the gel-plus-light and the membrane-plus-light
treated groups versus the control and light-only groups (collagen
orientation index, 0.18.+-.0.04, 0.21.+-.0.06 vs 0.50.+-.0.08,
0.52.+-.0.08. P<0.05). These results are also presented in a
graphical format in FIG. 6.
Based on the above findings from the in vivo dermal fibrotic
mouse--human skin graft model, these data indicate the potential
for the silicone-based biophotonic compositon of the present
disclosure for acceleration of wound healing and reduction of
fibrosis in human fibroproliferative disorders such as hypertrophic
scarring.
It should be appreciated that the invention is not limited to the
particular embodiments described and illustrated herein but
includes all modifications and variations falling within the scope
of the invention as defined in the appended claims.
* * * * *
References